Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Priority
Applicant’s statement “According to Electronic Priority Document Exchange (PDX) Program, the applicant submitted the application data sheet including WIPO Das code for the certified copy of foreign priority application, KR 10-2021-0135977 on August 29, 2022” filed on 11/10/2025 is acknowledged.
However, no English translation of Korean Patent Application No. 10-2021-013 5977 has been provided. Accordingly, the priority date of amended claim set filed on 08/29/2022 remains to be 06/17/2022, the filing date of PCT/KR2022/008633.
Should applicant desire to obtain the benefit of foreign priority under 35 U.S.C. 119(a)- (d) prior to declaration of an interference, a certified English translation of the foreign application must be submitted in reply to this action. 37 CFR 41.154(b) and 41.202(e).
Failure to provide a certified translation may result in no benefit being accorded for the non-English application.
Claim status
Applicant’s response filed on 11/10/2025 is acknowledged. Claims 1-3 are amended.
Claims 1-11 are pending. Claims 4-11 withdrawn from further consideration pursuant to 37 CFR 1.142(b), as being drawn to nonelected inventions, there being no allowable generic or linking claim. Claims 1-3 are currently under examination.
Summary of this Final Office Action
Previous rejection of claims 1-3 under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, is withdrawn as claims 1-3 are amended.
A new ground of rejection of claim 2 under 35 U.S.C. 102(b) necessitated by claim amendment filed on 11/10/2025 is documented in this Final office action.
A new ground of new matter rejection of claims 1-3 under 35 U.S.C. 112(a) necessitated by claim amendment filed on 11/10/2025 is documented in this Final office action.
Previous rejection of claim 1 under 35 U.S.C. 102(a)(1) as being anticipated by Thiebes et al. (2016) is withdrawn as claims 1-3 are amended.
A new ground of rejection of claim 1 under 35 U.S.C. 102(a)(1) necessitated by claim amendment filed on 11/10/2025 is documented in this Final office action.
Previous rejection of claims 1 and 2 under 35 U.S.C. 103 as being unpatentable over Thiebes et al. (2016) in view of Li et al. (2002) is withdrawn as claims 1-3 are amended.
A new ground of rejection of claims 1 and 2 under 35 U.S.C. 103 necessitated by claim amendment filed on 11/10/2025 is documented in this Final office action.
Previous rejection of claims 1 and 3 under 35 U.S.C. 103 as being unpatentable over Thiebes et al. (2016) in view of Weinandy et al. (2012) is withdrawn as claims 1-3 are amended.
A new ground of rejection of claims 1 and 3 under 35 U.S.C. 103 necessitated by claim amendment filed on 11/10/2025 is documented in this Final office action.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim 2 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Amended claim 2 filed on 11/10/2025 recites the limitation "the whole layer" in “wherein the whole layer of epithelial cells is formed by adhering a whole layer of the cultured epithelial cells to a planar support through electrostatic attraction”. There is insufficient antecedent basis for this limitation in the claim.
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1-3 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention. This is a new matter rejection.
Amended claim 1 filed on 11/10/2025 recites the limitation “wherein adjacent epithelial cells in the layer are directly contacting each other and are connected through tight junctions, adherens junctions, and desmosomes”, which does not have support from specification of instant application field on 08/29/2022
MPEP 2163 II at “(b) New Claims, Amended Claims, or Claims Asserting Entitlement to the Benefit of an Earlier Priority Date or Filing Date under 35 U.S.C. 119, 120, 365, or 386” states “To comply with the written description requirement of 35 U.S.C. 112(a) or pre-AIA 35 U.S.C. 112, first paragraph, or to be entitled to an earlier priority date or filing date under 35 U.S.C. 119, 120, 365, or 386, each claim limitation must be expressly, implicitly, or inherently supported in the originally filed disclosure.”
Applicant’s Arguments
Support for the amended language is found in the Specification as filed:
Paragraph [0027] discloses that:
"The epithelial cells are closely adjoined and are combined together through different cell
junctions, wherein each cell has polarity, and the surface of the epithelial cell may be
divided into an apical surface in contact with the lumen, a lateral surface in contact with
neighboring cells, and a bottom surface in contact with a basal lamina. "
This passage expressly describes that (i) the epithelial cells are closely adjoined (directly
contacting), (ii) they are combined together through different cell junctions, and (iii) each cell
exhibits polarity with an apical surface, lateral surface, and basal surface.
Furthermore, paragraph [0040] describes that culturing epithelial cells at an air-liquid
interface induces ciliation and confirms apical-basal orientation:
" ... the culturing of epithelial cells at the air-liquid interface may induce differentiation of
the epithelial cells into cilia cells, specifically, the cilia of epithelial cells contact air and the
bottom surface thereof may be cultured while being exposed to a culture medium. "
This establishes that the resulting structure has cilia on the apical surface facing the air
(lumen) and a basal surface in contact with the support.
A person of ordinary skill in the art ("POSITA") in epithelial biology would recognize that the phrase "different cell junctions" inherently refers to the well-known epithelial junctional
complex comprising tight junctions, adherens junctions, and desmosomes. See, e.g.:
• Alberts et al., Molecular Biology of the Cell (4th ed., 2002, Chapter 19): "Tight junctions occupy the most apical position in the junctional complex, followed by adherens junctions and desmosomes, which together provide strong mechanical coupling between adjacent epithelial cells." (attached hereto as EXHIBIT 1, printed from
<<https://www.ncbi.nlm.nih.gov/books/NBK26857/?utm_source=chatgpt.eom>>)
• Riibsam et al., Adherens Junctions and Desmosomes Coordinate Mechanics in
Epithelial Tissues, Front. Cell Dev. Biol. 6:76 (2018): "Cadherin-based adherens
junctions and desmosomes form, together with tight junctions, the epithelial intercellular
junctional complex that maintains cell-cell adhesion and epithelial integrity." (attached
hereto as EXHIBIT 2)
Thus, the amended claim merely enumerates the particular types of junctions encompassed by "different cell junctions" in paragraph [0027]. The recited polarity, cilia formation, and basal adhesion are likewise explicitly and inherently disclosed in paragraphs [0027] and [0040]. The Specification therefore conveys with reasonable clarity that the inventors were in possession of these specific structural features at the time of filing. See Ariad Pharm. v. Eli Lilly, 598 F.3d 1336 (Fed. Cir. 2010) (en bane).
Response to Applicant’s Arguments
The examiner acknowledged that how “epithelial cells contacting each other and are connected through tight junctions, adherens junctions, and desmosomes” are textbook level knowledge for a skilled artisan. However, this does not in any way collaborate with Applicant’s arguments that the specification of instant application field on 08/29/2022 could support the notion that Applicants have the possession of claimed products because the generic statements disclosed in paragraphs [0027] and [0040] of instant specification regarding “cell junctions” (a genus limitation) does not inherently render Applicant’s possession of “tight junctions, adherens junctions, and desmosomes” (a species limitation) recited in amended claim 1 filed on 11/10/2025.
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
Claim 1 is rejected under 35 U.S.C. 102(a)(1) as being anticipated by Thiebes et al. (2016) (Thiebes et al. Flexible Endoscopic Spray Application of Respiratory Epithelial Cells as Platform Technology to Apply Cells in Tubular Organs, Tissue Eng Part C Methods, 2016 Apr;22(4):322-31. doi: 10.1089/ten.TEC.2015.0381. Epub 2016 Mar 18) as evidenced by Ojakian et al. (2001) (Ojakian et al. Integrin regulation of cell-cell adhesion during epithelial tubule formation, J Cell Sci., 2001 Mar;114(Pt 5):941-52. doi: 10.1242/jcs.114.5.941).
Amended claim 1 filed on 11/10/2025 reads as follows: An epithelial cell tube, comprising: a columnar support; and a layer of epithelial cells positioned on an outer peripheral surface of the columnar support, wherein the epithelial cells are oriented so that cilia of the epithelial cells face a lumen side, wherein adjacent epithelial cells in the layer are directly contacting each other and are connected through tight junctions, adherens junctions, and desmosomes, and wherein the epithelial cells exhibit a polarized morphology having cilia formed on an apical surface facing the lumen side and a basal surface adhered to the columnar support.
Claim interpretations: The limitation “wherein adjacent epithelial cells in the layer are directly contacting each other and are connected through tight junctions, adherens junctions, and desmosomes” is the intrinsic characteristics of epithelial cell junctions, which is evidenced by Ojakian et al. (2001) further discussed below.
Regarding claim 1, Thiebes et al. (2016) teaches that “We designed a flexible endoscopic spraying device based on a bronchoscope and tested it with respiratory epithelial cells. With this device cells can also be applied in a thin layer of fibrin glue. We evaluated the survival rate directly after spray application with a live-dead staining and the long-term differentiation
capacity with histology and electron microscopy. Furthermore, the random distribution of cells when applied in a tube was analyzed and the macroscopic and microscopic characteristics of the endoscopic spray were investigated using high-speed visualization.” (See Method). Thiebes et al. (2016) teaches that “Spray visualization revealed a polydisperse character of the spray with the majority of droplets larger than epithelial cells. Spray application does not influence the survival rate and differentiation of respiratory epithelial cells. After 4 weeks, cells built up a pseudostratified epithelial layer with cilia and goblet cells. When cells are applied in a thin layer of fibrin gel into a tube, a nearest neighbor index of 1.2 is obtained, which suggests a random distribution of the cells.” (See Results).
Regarding the limitation “wherein the epithelial cells are oriented so that cilia of the epithelial cells face a lumen side” and the limitation “wherein the epithelial cells exhibit a polarized morphology having cilia formed on an apical surface facing the lumen side and a basal surface adhered to the columnar support” recited in claim 1, Thiebes et al. (2016) teaches “Thus, as new method for palliative treatment of patients with malignant airway stenosis we developed the PulmoStent concept, based on the vascular BioStent, as shown in Figure 1. When combining a covered metal stent with a tissue engineered epithelial lining, mucostasis and the migration risk can be overcome.” (See right column, page 322),
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FIG. 1. PulmoStent concept. A metal stent with a nutrientpermeable cover is coated with respiratory epithelial cells for mucociliary clearance. Color images available online at www.liebertpub.com/tec
Moreover, regarding the limitation “a layer of epithelial cells positioned on an outer peripheral surface of a columnar support” recited in claim 1, Thiebes et al. (2016) teaches that “Figure 7A–F depicts sections stained by PAS, Figure 7G–I shows scanning electron microscopy images. In all groups, a pseudostratified appearance with columnar-shaped cells is seen with basal cells on the lower side. Apically, there are ciliated cells (arrows) and goblet cells (arrowheads) present. The cilia can also be seen in the electron microscopy images. Figure 7J–O shows immunohistochemistry stainings of the respective groups. Figure 7J–L shows uniform pancytokeratin expression in all groups. Figure 7M–O shows uniform expression of Claudin-1. Claudin-1 is a protein of tight junctions that proves the barrier function of the epithelial layer.” (See right column, page 326).
Thiebes et al. (2016) furtehr teaches under “Spraying in tubes section” that “To find out if the cells are distributed randomly when coating the lumen of a tube, a nearest neighbor analysis was conducted. The lumens of three 20 mL syringes (inner diameter: 20 mm; BD Medical) were covered with a piece of paper, which was subsequently coated with cells by slowly moving the bronchoscope tip inside the syringe. We used vascular smooth muscle cells that were stained with calcein AM before spraying (isolation and culture was described by Tschoeke et al.).” (See bridging paragraph, page 324-325). It is noted that “the cells are distributed randomly” taught by Thiebes et al. (2016) refers to random distribution of epithelial cells during spraying in tubes onto a catheter/stent [See Fig, 2 Spraying setup and Fig. 4 of Thiebes et al. (2016)], not random orientation of epithelial cells positioned on an outer peripheral surface of a columnar support.
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FIG. 7. Long-term differentiation of respiratory epithelial cells. Nonsprayed control (A, D, G, J, M), cells after spray application without (B, E, H, K, N) and with fibrin gel (C, F, I, L, O). (A–F) Periodic acid Schiff’s reaction; (D–F) show magnifications of (A–C). Epithelial cells show a columnar shape with cilia on all samples (arrows); goblet cells can also be seen (arrowheads). (G–I) Scanning electron microscopy images show cilia on cell surface in all
samples. (J–O) Immunohistochemistry of pancytokeratin (J–L) and claudin- 1 (M–O) show uniform expression of the epithelial marker and tight junction proteins, respectively. Color images available online at www.liebertpub.com/tec
Regarding the limitation “wherein adjacent epithelial cells in the layer are directly contacting each other and are connected through tight junctions, adherens junctions, and desmosomes”, Ojakian et al. (2001) teaches that “The extracellular matrix plays an important role in regulation of epithelial development and organization. To determine more precisely the function of extracellular matrix in this process, the initial steps in collagen-mediated formation of epithelial tubules were studied using a model cell culture system. Previous studies have demonstrated that incubation of Madin-Darby canine kidney (MDCK) epithelial cells with a collagen gel overlay induces (beta)1 integrin-regulated epithelial remodeling accompanied by extensive cell rearrangements and formation of epithelial tubules. During epithelial remodeling there was extensive disruption of the epithelial junctional complex. Progressive opening of tight junctions was observed over 8 hours using transepithelial resistance measurements and immunofluorescence microscopy demonstrated that tight and adherens junction proteins were dispersed throughout the apical and basolateral membranes. Junction complex disruption allowed the formation of apical cell extensions and subsequent migration of selected cell sheets from the epithelial monolayer. Confocal microscopy demonstrated the presence of adherens junction (E-cadherin, (alpha)-catenin, (beta)-catenin, plakoglobin) and desmosomal (desmoplakin-1/2, plakoglobin) proteins on, and within, cell extensions demonstrating that cell junctions had undergone considerable disassembly. However, groups of cell extensions appeared to be associated by E-cadherin/catenin-mediated interactions. Association of E-cadherin/catenin complexes with the epithelial cytoskeleton was analyzed by differential detergent extraction. SDS-PAGE and immunoblot analysis demonstrated that adherens junction proteins were primarily cytoskeleton-associated in control cells. During integrin-regulated remodeling, there was a progressive reduction in the interaction of adherens junction proteins with the cytoskeleton suggesting that they play an important role in the maintenance of epithelial integrity. Since loss of transepithelial electrical resistance and disruption of junctional complexes were inhibited by an antifunctional integrin antibody, we propose that activation of integrin signaling pathways regulate junctional complex stability, cell-cell interactions and cell migration. These observations provide evidence that integrin-regulated MDCK epithelial tubule formation can serve as a model system for studying rearrangements of epithelial sheets which occur during development (See Abstract).
Applicant’s Arguments
A claim is anticipated under 35 U.S.C. § 102 only if a single prior-art reference expressly
or inherently discloses each and every element of the claimed invention, arranged as in the claim.
See Verdegaal Bros., Inc. v. Union Oil Co., 814 F.2d 628, 631 (Fed. Cir. 1987); In re Gleave, 560 F.3d 1331, 1334 (Fed. Cir. 2009). Anticipation is not established by showing that the prior art is merely capable of being modified to meet the claim, or that it achieves a similar result. Each claimed feature must be found in the same structure, as arranged and functioning together.
Response to Applicant’s Arguments
As documented above, the amended claim 1 filed on 11/10/2025 is rejected under 35 U.S.C. 102(a)(1) as being anticipated by Thiebes et al. (2016) (Thiebes et al. Flexible Endoscopic Spray Application of Respiratory Epithelial Cells as Platform Technology to Apply Cells in Tubular Organs, Tissue Eng Part C Methods, 2016 Apr;22(4):322-31. doi: 10.1089/ten.TEC.2015.0381. Epub 2016 Mar 18) as evidenced by Ojakian et al. (2001) (Ojakian et al. Integrin regulation of cell-cell adhesion during epithelial tubule formation, J Cell Sci., 2001 Mar;114(Pt 5):941-52. doi: 10.1242/jcs.114.5.941.
The limitation “wherein adjacent epithelial cells in the layer are directly contacting each other and are connected through tight junctions, adherens junctions, and desmosomes” is the intrinsic characteristics of epithelial cell junctions, which is evidenced by the teachings of Ojakian et al. (2001).
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claims 1 and 2 are rejected under 35 U.S.C. 103 as being unpatentable over Thiebes et al. (2016) in view of Ojakian et al. (2001) (Ojakian et al. Integrin regulation of cell-cell adhesion during epithelial tubule formation, J Cell Sci., 2001 Mar;114(Pt 5):941-52. doi: 10.1242/jcs.114.5.941), and Li et al. (2002) (Li et al., Using microfabrication and electrostatic layer-by-Layer (LbL) self-assembly technologies to improve the growth and alignment of smooth muscle cells, 2nd Annual International IEEE-EMBS Special Topic Conference on …, 2002• ieeexplore.ieee.org).
Amended Claim 2 filed on 11/10/2025 reads as follows: The epithelial cell tube according to claim 1, wherein the whole layer of epithelial cells is formed by adhering a whole layer of the cultured epithelial cells to a planar support through electrostatic attraction.”.
The teachings of Thiebes et al. (2016) have been documented above in the rejection of Claim 1 under 35 U.S.C. 102(a)(1).
Regarding the limitation “wherein adjacent epithelial cells in the layer are directly contacting each other and are connected through tight junctions, adherens junctions, and desmosomes”, Ojakian et al. (2001) teaches that “The extracellular matrix plays an important role in regulation of epithelial development and organization. To determine more precisely the function of extracellular matrix in this process, the initial steps in collagen-mediated formation of epithelial tubules were studied using a model cell culture system. Previous studies have demonstrated that incubation of Madin-Darby canine kidney (MDCK) epithelial cells with a collagen gel overlay induces (beta)1 integrin-regulated epithelial remodeling accompanied by extensive cell rearrangements and formation of epithelial tubules. During epithelial remodeling there was extensive disruption of the epithelial junctional complex. Progressive opening of tight junctions was observed over 8 hours using transepithelial resistance measurements and immunofluorescence microscopy demonstrated that tight and adherens junction proteins were dispersed throughout the apical and basolateral membranes. Junction complex disruption allowed the formation of apical cell extensions and subsequent migration of selected cell sheets from the epithelial monolayer. Confocal microscopy demonstrated the presence of adherens junction (E-cadherin, (alpha)-catenin, (beta)-catenin, plakoglobin) and desmosomal (desmoplakin-1/2, plakoglobin) proteins on, and within, cell extensions demonstrating that cell junctions had undergone considerable disassembly. However, groups of cell extensions appeared to be associated by E-cadherin/catenin-mediated interactions. Association of E-cadherin/catenin complexes with the epithelial cytoskeleton was analyzed by differential detergent extraction. SDS-PAGE and immunoblot analysis demonstrated that adherens junction proteins were primarily cytoskeleton-associated in control cells. During integrin-regulated remodeling, there was a progressive reduction in the interaction of adherens junction proteins with the cytoskeleton suggesting that they play an important role in the maintenance of epithelial integrity. Since loss of transepithelial electrical resistance and disruption of junctional complexes were inhibited by an antifunctional integrin antibody, we propose that activation of integrin signaling pathways regulate junctional complex stability, cell-cell interactions and cell migration. These observations provide evidence that integrin-regulated MDCK epithelial tubule formation can serve as a model system for studying rearrangements of epithelial sheets which occur during development (See Abstract).
Thiebes et al. (2016) and Ojakian et al. (2001) do not explicitly teach the limitation “the whole layer of epithelial cells is formed by adhering a whole layer of the cultured epithelial cells to a planar support through electrostatic attraction” recited in claim 2.
Regarding the limitation “adhering a whole layer of the cultured epithelial cells to a planar support through electrostatic attraction” recited in claim 2, Li et al. (2002) teaches “Smooth muscle cells (SMCs) were cultured on polydimethylsiloxane (PDMS) based cell culture substrates. Two types of experiments were performed to address the cell behaviors on these substrates. One was culturing smooth muscle cells on bare PDMS flat surfaces and gelatin coated PDMS flat surfaces deposited using electrostatic layer-by-layer self-assembly technology. The other was culturing smooth muscle cells on two micro-structured PDMS microchannel substrates and PDMS flat surface substrates. The microchannels are 5-mm channels (line width = 5 mm, spacing width = 5 mm, depth = 5 mm) and 100-mm channels (line width = 100 mm, spacing width = 100 mm, depth = 50 mm) respectively. All substrates were coated with multilayers (50 nm in thickness) of gelatin using electrostatic layer-by-layer self-assembly technology in order to improve the attachment of the cells. We concluded that surface treatment, such as gelatin coating, is able to help smooth muscle cells attach on PDMS substrates. Accordingly, it will increase the potential growth of cells on the engineered PDMS substrates. Second, smooth muscle cells showed a clear preference of alignment long the channel sidewall on the 100-mm channel substrate as compared to that on the flat surface substrate. Microchannels are able to align the growth of smooth muscle cells, and the ability of controlling the alignment depends on the dimension of the microstructures, as well as the surface treatment for increasing cell attachment. Microfabrication and electrostatic layer-by-layer self-assembly technologies have significant potential for application in the field of tissue engineering.” (See Abstract).
Moreover, Li et al. (2002) teaches that PolyStyreneSulfonate (PSS) and gelatin are negatively charged. PolyDimethylDiallylAmmonium chloride (PDDA) is positively charged; and PolyDiMethylSiloxane (PDMS) substrate carrying a negative surface charge (See left column, 2nd page, under Electrostatic layer-by-layer self-assembly section). Li et al. (2002) further teaches that “In our experiments, we compared the number of cells growing on the bare PDMS flat surface and that on the gelatin coated PDMS flat surface. We also measured the alignment angles on the flat PDMS surface substrate and the 100-μm channel PDMS substrate. From these collected data, we investigated the attachment and spreading of smooth muscle cells on the microfabricated substrates as well as the biocompatibility of the materials after surface modifications. Both of the methods are based on optical microscopy images collected during cell
culture (See right column, 2nd page, under Data Measurement and Statistical Analysis section).
It would have been prima facie obvious for a skilled artisan to incorporate the teachings of Li et al. (2002) into the combined teachings of Thiebes et al. (2016) and Ojakian et al. (2001) because Li et al. (2002) provides detailed information regarding using microfabrication and electrostatic layer-by-Layer (LbL) self-assembly technologies to improve the growth and alignment of smooth muscle cells.
There would have been a reasonable expectation of success for a skilled artisan to incorporate the teachings of Li et al. (2002) into the combined teachings of Thiebes et al. (2016) and Ojakian et al. (2001) because Li et al. (2002) specifically teaches that “After coating gelatin on the PDMS surface, cells showed a preference to grow and propagate on the gelatin coated flat surface” (See left column, 4th page, under “B. Cell Culture Results” section, starting on the right column, of the 3rd page); and that “Since smooth muscle cells could not grow on 5-mm channel substrate, we also cultured the cells on a 100-mm channel substrate. Figure 5 are the images of smooth muscle cells cultured on the 100-mm channel substrate. We found that smooth muscle cells not only grew on this type substrate, but also showed a clear preference for alignment along the channel sidewall on the 100-µm channel substrate as compared to on the flat surface substrate (See right column, 5th page).
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Figure 5. Optical microscopy images of smooth muscle cells cultured on 100-μm channel substrate, indicating that cells could not only grow on the channel surface, but also showing a clear preference of alignment along the channel sidewall.
Applicant’s Arguments
Thiebes discloses spray deposition of epithelial cells suspended in fibrin glue onto a
tubular scaffold, yielding a random distribution of isolated cells (nearest-neighbor index = 1.2).
Such distribution is the antithesis of a confluent epithelium with continuous junctional
complexes. Specifically, Thiebes fails to disclose: Cells in direct contact forming tight, adherens, and desmosomal junctions; A polarized morphology with apical cilia and basal adhesion; or
Any culturing or differentiation under air-liquid interface (ALI) conditions that could yield such structure.
Li et al. (2002) describes planar transwell membranes fabricated by electrostatic layer-by-layer (LbL) assembly for culturing smooth muscle cells, not epithelial cells. Li's disclosure is limited to two-dimensional planar systems, not tubular scaffolds.
Importantly, Li's LbL technology addresses cell alignment and substrate charge control,
not epithelial polarity or tight-junction formation.
Li contains no teaching or suggestion of forming a confluent epithelial layer at an air-liquid interface on a three-dimensional tube.
Moreover, Li's planar culture conditions are biologically and mechanically incompatible with Thiebes's fibrin-based coating process.
As described in the Specification, ALL culture on a tubular support yields tight junctional
continuity, uniform ciliation, and lumen-facing polarity, none of which are possible in the
submerged or planar systems of Thiebes or Li.
Such results are unexpected and constitute strong objective evidence of nonobviousness.
See In re Soni, 54 F.3d 746 (Fed. Cir. 1995).
Response to Applicant’s Arguments
The limitation “wherein adjacent epithelial cells in the layer are directly contacting each other and are connected through tight junctions, adherens junctions, and desmosomes” is taught by Ojakian et al. Integrin regulation of cell-cell adhesion during epithelial tubule formation, J Cell Sci., 2001 Mar;114(Pt 5):941-52. doi: 10.1242/jcs.114.5.941).
The isolation and cultural conditions of epithelial cells have been explicitly taught by both Thiebes et al. (2016) (page 323) and Ojakian et al. (2001) (page 942). Culturing epithelial cells and coating epithelial cells onto desired columnar support are two separate steps, and there is nothing “biologically and mechanically incompatible” as explicitly demonstrated by Thiebes et al. (2016). Furthermore, there is no requirement of “forming a confluent epithelial layer at an air-liquid interface on a three-dimensional tube” recited in amended claim 2 as Applicant argued. Moreover, Li et al., is not relied on for the teachings of “epithelial polarity or tight-junction formation” which is prima facie obvious based on the combined teachings of Thiebes et al. (2016) and Ojakian et al. (2001). Rather, Li et al. (2002) is relied on for the teachings regarding the limitation “adhering a whole layer of the cultured epithelial cells to a planar support through electrostatic attraction” recited in claim 2, and the teachings by Li et al. (2002) provide the molecular interactions in coating or culturing a layer of epithelial cells onto a desired columnar support taught by Thiebes et al. (2016) and Ojakian et al. (2001).
In response to applicant's arguments against the references individually, one cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981); In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986).
The unexpected results regarding “culture on a tubular support yields tight junctional continuity, uniform ciliation, and lumen-facing polarity” are not persuasive based on the combined teachings of Thiebes et al. (2016) (Thiebes et al. Flexible Endoscopic Spray Application of Respiratory Epithelial Cells as Platform Technology to Apply Cells in Tubular Organs, Tissue Eng Part C Methods, 2016 Apr;22(4):322-31) and Ojakian et al. (2001) (Ojakian et al. Integrin regulation of cell-cell adhesion during epithelial tubule formation, J Cell Sci., 2001 Mar;114(Pt 5):941-52. doi: 10.1242/jcs.114.5.941) documented in this Final office action.
Claims 1 and 3 are rejected under 35 U.S.C. 103 as being unpatentable over Thiebes et al. (2016) in view of Ojakian et al. (2001) (Ojakian et al. Integrin regulation of cell-cell adhesion during epithelial tubule formation, J Cell Sci., 2001 Mar;114(Pt 5):941-52. doi: 10.1242/jcs.114.5.941), and Weinandy et al. (2012) (Weinandy et al., The BioStent: Novel Concept for a Viable Stent Structure, Tissue Eng Part A, 2012 Sep;18(17-18):1818-26. doi: 10.1089/ten. TEA.2011.0648. Epub 2012 Jun 7).
Amended claim 3 filed on 11/10/2025 reads as follows: The epithelial cell tube according to claim 1, wherein the tube is formed by rolling a planar support carrying the layer of epithelial cells into a tubular shape and linking both ends of the planar support together, such that the layer of epithelial cells faces outward on the outer peripheral surface of the columnar support.
The teachings of Thiebes et al. (2016) have been documented above in the rejection of claim 1 under 35 U.S.C. 102(a)(1).
MPEP 2113 Product-by-Process Claims [R-01.2024]
PRODUCT-BY-PROCESS CLAIMS ARE NOT LIMITED TO THE MANIPULATIONS OF THE RECITED STEPS, ONLY THE STRUCTURE IMPLIED BY THE STEPS
"[E]ven though product-by-process claims are limited by and defined by the process, determination of patentability is based on the product itself. The patentability of a product does not depend on its method of production. If the product in the product-by-process claim is the same as or obvious from a product of the prior art, the claim is unpatentable even though the prior product was made by a different process." In re Thorpe, 777 F.2d 695, 698, 227 USPQ 964, 966 (Fed. Cir. 1985) (citations omitted) (Claim was directed to a novolac color developer).
Regarding the limitation “wherein adjacent epithelial cells in the layer are directly contacting each other and are connected through tight junctions, adherens junctions, and desmosomes”, Ojakian et al. (2001) teaches that “The extracellular matrix plays an important role in regulation of epithelial development and organization. To determine more precisely the function of extracellular matrix in this process, the initial steps in collagen-mediated formation of epithelial tubules were studied using a model cell culture system. Previous studies have demonstrated that incubation of Madin-Darby canine kidney (MDCK) epithelial cells with a collagen gel overlay induces (beta)1 integrin-regulated epithelial remodeling accompanied by extensive cell rearrangements and formation of epithelial tubules. During epithelial remodeling there was extensive disruption of the epithelial junctional complex. Progressive opening of tight junctions was observed over 8 hours using transepithelial resistance measurements and immunofluorescence microscopy demonstrated that tight and adherens junction proteins were dispersed throughout the apical and basolateral membranes. Junction complex disruption allowed the formation of apical cell extensions and subsequent migration of selected cell sheets from the epithelial monolayer. Confocal microscopy demonstrated the presence of adherens junction (E-cadherin, (alpha)-catenin, (beta)-catenin, plakoglobin) and desmosomal (desmoplakin-1/2, plakoglobin) proteins on, and within, cell extensions demonstrating that cell junctions had undergone considerable disassembly. However, groups of cell extensions appeared to be associated by E-cadherin/catenin-mediated interactions. Association of E-cadherin/catenin complexes with the epithelial cytoskeleton was analyzed by differential detergent extraction. SDS-PAGE and immunoblot analysis demonstrated that adherens junction proteins were primarily cytoskeleton-associated in control cells. During integrin-regulated remodeling, there was a progressive reduction in the interaction of adherens junction proteins with the cytoskeleton suggesting that they play an important role in the maintenance of epithelial integrity. Since loss of transepithelial electrical resistance and disruption of junctional complexes were inhibited by an antifunctional integrin antibody, we propose that activation of integrin signaling pathways regulate junctional complex stability, cell-cell interactions and cell migration. These observations provide evidence that integrin-regulated MDCK epithelial tubule formation can serve as a model system for studying rearrangements of epithelial sheets which occur during development (See Abstract).
Thiebes et al. (2016) and Ojakian et al. (2001) does not explicitly teach the limitation “wherein the tube is formed by rolling a planar support carrying the layer of epithelial cells into a tubular shape and linking both ends of the planar support together, such that the layer of epithelial cells faces outward on the outer peripheral surface of the columnar support” recited in amended claim 3.
Regarding the limitation “wherein the tube is formed by rolling a planar support carrying the layer of epithelial cells into a tubular shape and linking both ends of the planar support together, such that the layer of epithelial cells faces outward on the outer peripheral surface of the columnar support” recited in amended claim 3, Weinandy et al. (2012) teaches that “Materials and Methods: Small-caliber BioStents (Ø= 6 mm; n = 4) were produced by casting a nitinol stent within a thin fibrin/vascular smooth muscle cell (vSMC) mixture, followed by luminal endothelial cell seeding, and conditioning of the BioStent within a bioreactor system. The potential remodeling of the fibrin component into tissue was analyzed using routine histological methods. Scanning electron microscopy was used to assess the luminal endothelial cell coverage following the conditioning phase and crimping of the stent. Results: The BioStent was shown to be noncytotoxic, with no significant effect on cell proliferation. Gross and microscopic analysis revealed complete integration of the nitinol component within a viable tissue structure. Hematoxylin and eosin staining revealed a homogenous distribution of vSMCs throughout the thickness of the BioStent, while a smooth, confluent luminal endothelial cell lining was evident and not significantly affected by the crimping/release process.” (See page 1818).
Weinandy et al. (2012) teaches that “In the present study, we propose a novel approach to address this biocompatibility issue by ensheathing a warp-knitted, self-expanding nitinol stent within a cell-seeded fibrin matrix (termed BioStent) (Fig. 1) (See left column, page 1819).
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FIG. 1. Illustration of the BioStent concept. (1) The foreign body reaction, (2) the cell proliferation with ingrowth into the lumen, and (3) acute thrombosis by hemo-incompatibility
need to be avoided in stent angioplasty.
Weinandy et al. (2012) further teaches that “To obtain stent structures with sufficient radial stability, superelastic nitinol wire (NiTi Nr.1SE; Fort Wayne Metals) was employed (76-mm thickness). To achieve adequate surface area coverage, warp-knitted stents with a two-bar tricot interloop were produced on a double-raschel warp-knitting machine (Karl Mayer). With regard to loop density, 12 loops/cm were produced with a needle gauge of 30 needles/inch (E30) (Fig. 2). The fabricated tubular nitinol structures were fixed on a metal core and tempered at 5000C for 15 min in a high-temperature kiln (Heraeus) to achieve the superelastic phase.” (See bridging paragraph, page 1819-1820).
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FIG. 2. (A) CAD graph of the stent structure with a two-bar tricot interloop); (B)
warp-knitted nitinol stent. CAD, computer-aided design.
It would have been prima facie obvious for a skilled artisan to incorporate the teachings of Weinandy et al. (2012) into the combined teachings of Thiebes et al. (2016) and Ojakian et al. (2001) because Weinandy et al. (2012) provides the teachings regarding “The BioStent: Novel Concept for a Viable Stent Structure” (See Title, page 1818); and “The concept combines the principles of vascular tissue engineering with a self-expanding stent: casting of the stent within a cellularized fibrin gel structure, followed by bioreactor conditioning, allows complete integration of the stent within engineered tissue.” (See Design, page 1818).
There would have been a reasonable expectation of success for a skilled artisan to incorporate the teachings of Weinandy et al. (2012) into the combined teachings of Thiebes et al. (2016) and Ojakian et al. (2001) because Weinandy et al. (2012) specifically teaches that “To achieve adequate surface area coverage, warp-knitted stents with a two-bar tricot interloop were produced on a double-raschel warp-knitting machine (Karl Mayer). With regard to loop density, 12 loops/cm were produced with a needle gauge of 30 needles/ inch (E30) (Fig. 2).”
Applicant’s Arguments
Weinandy is directed to mechanical stent and endoprosthesis design, emphasizing metallic or polymeric meshes with controlled radial expansion and flexibility. Its "linking" refers to mechanical joining of rigid struts or ends-through welding, crimping, or thermal bonding-to form a tubular structure. Weinandy nowhere addresses biological coating, epithelial differentiation, or ALI culture. Its disclosure is therefore non-analogous art (In re Clay, 966 F.2d 656 (Fed. Cir. 1992)), as it pertains to solving an entirely different problem-mechanical durability rather than biological integrity. In contrast, amended claim 1 recites a biological epithelial construct defined by junctional, ciliary, and polarity structures, which Weinandy neither discloses nor suggests.
Response to Applicant’s Arguments
Amended claim 3 filed on 11/10/2025 reads as follows: The epithelial cell tube according to claim 1, wherein the tube is formed by rolling a planar support carrying the layer of epithelial cells into a tubular shape and linking both ends of the planar support together, such that the layer of epithelial cells faces outward on the outer peripheral surface of the columnar support.
Claim 3 as written is a product-by-process claim and determination of patentability is based on the product itself.
MPEP 2113 Product-by-Process Claims [R-01.2024]
PRODUCT-BY-PROCESS CLAIMS ARE NOT LIMITED TO THE MANIPULATIONS OF THE RECITED STEPS, ONLY THE STRUCTURE IMPLIED BY THE STEPS
"[E]ven though product-by-process claims are limited by and defined by the process, determination of patentability is based on the product itself. The patentability of a product does not depend on its method of production. If the product in the product-by-process claim is the same as or obvious from a product of the prior art, the claim is unpatentable even though the prior product was made by a different process." In re Thorpe, 777 F.2d 695, 698, 227 USPQ 964, 966 (Fed. Cir. 1985) (citations omitted) (Claim was directed to a novolac color developer).
Applicant’s argument that “Weinandy nowhere addresses biological coating, epithelial differentiation, or ALL culture. Its disclosure is therefore non-analogous art” is not persuasive because the teachings by Weinandy is directed to “The concept combines the principles of vascular tissue engineering with a self-expanding stent” (See Fig. 1 and Fig. 2 of Weinandy), which is clearly analogous art in the same field of endeavor of “Flexible Endoscopic Spray Application of Respiratory Epithelial Cells as Platform Technology to Apply Cells in Tubular Organs” taught by Thiebes et al. (2016) (See Fig. 1 and Fig. 2 of Thiebes et al.).
In response to applicant's arguments against the references individually, one cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981); In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986).
Conclusion
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/WU CHENG W SHEN/Supervisory Patent Examiner, Art Unit 1682