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 .
DETAILED ACTION
This action is in response to papers filed April 3, 2025. Claims 1-14, 16-18 and 20-21 are currently pending. It is noted that claims 1, 14, 17, and 20 are independent claims.
Therefore, claims 1-14, 16-18 and 20-21 are under examination to which the following grounds of rejection are applicable.
Specification
The listing of references in the specification is not a proper information disclosure statement. 37 CFR 1.98(b) requires a list of all patents, publications, or other information submitted for consideration by the Office, and MPEP § 609.04(a) states, "the list may not be incorporated into the specification but must be submitted in a separate paper." Therefore, unless the references have been cited by the examiner on form PTO-892, they have not been considered.
Priority
The instant application claims priority to International Application PCT/US2023/062581 filed on February 14, 2023. The International Application claims priority to Provisional Application 63/310,365 filed February 15, 2022.
Therefore, the earliest effective filing date for the instant application is February 15, 2022.
Claim Objections
Claim 1 and 8 are objected to because of the following informalities:
Claim 1 and 8 lack the proper definitive article providing proper antecedent basis for the phrase “culturing NFBs” in line 4. The claim should be amended to state “culturing the NFBs”.
Appropriate correction is required.
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.
Claims 4-5 and 16 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.
Regarding claim 4, it is indefinite in its recitation of the term “nerve NFBs” in line 1. There is not proper antecedent basis in the claim, as claim 1 only recites “NFBs”.
Regarding claim 16, the term “significant” is a relative term which renders the claim indefinite. The term “significant amount” is not defined by the claim, the specification does not provide a standard for ascertaining the requisite degree, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention. Regarding claim 16, it is indefinite in its recitation of the term “the progenitor cell” in line 1 as it lacks proper antecedent basis. Claim 14 recites “a human neuronal progenitor cell”.
Claim 5 is rendered indefinite insofar that it depends on claim 4.
Claim Rejections - 35 USC § 112: Scope of Enablement
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 1-14, 16-18 and 20-21 rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for
Methods of using subcutaneous adipose tissue comprising nerve fibre bundles (NFBs) comprising neural crest (NC)-derived Schwann cells;
does not reasonably provide enablement for method of generation of human neuronal progenitor cells with NFBs without NC-derived Schwann cells. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the invention commensurate in scope with these claims.
The criteria for enablement set out In re Wands, MPEP 2162.01(a) , considers the following factors:
Breadth of the Claims
The claims are broadly directed to a method of producing neural progenitor cells from any source of adipose tissue to collect nerve fiber bundles (NFBs) and culturing NFBs to generate human neuronal progenitor cells. The claims are broadly but reasonably interpreted as generating any human neuronal progenitor cells from collected any dissociated adipose tissue comprising NFBs but that do not necessary comprise neural crest (NC)-derived Schwann cells. As such, the breadth of the claims is directed to a genus of dissociated adipose tissues comprising nerve fibre bundles and producing human neuronal progenitor cells. The claims are only enabling for subcutaneous adipose tissue comprising NFBs comprising neural crest (NC)-derived Schwann cells as the source of human neuronal progenitor cells from adipose tissue.
State of Prior Art
Nagase et al. (Published: 2008. Cited in IDS filed 4/03/2025. WO2008150001A1. Machine Translation provided by Examiner) teaches that neural stem cells are derived from adipose tissue (para 0130, “, the cells derived from adipose tissue…While not being bound by theory, one can also consider the advantage of using cells with pluripotency that can easily induce Musashi-positive (or nestin-positive) cells, kn While not being bound by theory, one can also consider the advantage of using cells with pluripotency that can easily induce Musashi-positive (or nestin-positive) cells, known as markers of neural stem cells .own as markers of neural stem cells.”) Moreover, Nagase teaches that the adipose tissue-derived cells were harvested from dissociating the tissue and culturing the pellets (para 0020, step 5, “The adipose tissue-derived cells are either stromal-vascular-fraction cells from which adipose tissue is harvested, collagenase treatment, and then centrifuged and collected as pellets, or adipose-derived stem cells derived from waste fluid, according to the above items.”). As such, the art is silent on whether stem cells can be isolated from adipose tissue comprising nerve fiber bundles without Schwann cells.
Lee et al. (Published 2021. International journal of molecular sciences, 22(8), 4141.) discusses isolated peripheral-nerve derived stem cells (PNSCs) from adult peripheral nerves (Abstract, “Here, we isolated novel peripheral nerve-derived stem cells (PNSCs) from adult peripheral nerves with similar characteristics to neural-crest stem cells.”). Moreover, Lee teaches obtaining PNSCs from peripheral nerves by dissociating nerve segments in collagenase to obtain PNSCs (pp. 14, para 4, “After 14 days of organ culture, collagen hydrogels containing PN fragments and outgrown PNSCs were incubated with 0.01% collagenase type I at 37 °C under dynamic conditions in an orbital shaker at 25 rpm for 30 min”).
The art is silent on whether culturing nerve fiber bundles (i.e. the axon) alone will successfully result in the isolation of human neuronal progenitor cells. Therefore, the method of isolating neuronal progenitor cells from nerve fiber bundles alone is very unpredictable in the art, and a skilled artisan would not expect that culturing NFBs alone would result in the production of neuronal progenitor cells without undue experimentation.
Amount of Direction Provided by the Inventor
The Specification provides insufficient direction and guidance to enable a person of ordinary skill in the art to practice the full scope of the claimed invention, as the Specification only discloses a Schwann cell-associated example rather than the full scope of the claims. Although claim 1 broadly encompasses any human neural progenitor cell population by culturing adipose tissue-derived nerve fiber bundles, the Specification only teaches that neurogenic potential can be found in Schwann cells derived from adipose tissue, specifically that they are derived from the nerve fiber bundles within adipose tissue (para 0073, “ However, no equivalent [neural] progenitors are known to exist postnatally in the nerve fiber niche. It is now shown that neural crest (NC)-derived Schwann cells isolated from the adipose tissue, e.g., subcutaneous adipose tissue (SAT) or visceral adipose tissue (VAT), are the only cell population displaying gliogenic and neurogenic potential consistent with adipose tissue-derived neural stem cells (NSCs), e.g., SAT-derived NSCs (SAT-NSCs) or VAT-derived NSCs (VAT-NSCs)”). Moreover, the working examples conclude that “NC-derived Schwann cells are able to give rise to SAT-NSCs” (para 0102, Example 1), thereby indicating that neuronal stem cells (NSCs) can be isolated from Schwann cells rather than the nerve fiber bundles themselves. Accordingly, the Specification only provides guidance through the disclosed Schwann cell-associated pathway and does not teach how to produce neuronal progenitor cells across the full breadth of the claims.
Presence or Absence of Working Examples
The Specification only provides working examples for deriving stem cells from NC-derived Schwann cells that reside within nerve fiber bundles, and that Wnt1 positive cells were derived from the NFBs (para 0100, “ Neural Crest (NC)-Derived Schwann Cells Reside in NFBs Coursing Through the SAT [subcutaneous adipose tissue]”; para 0101, “Filtered cells, which excludes the NFBs, from the SAT cultured in low attachment conditions with neuroproliferation medium formed spheroids in vitro and increased the proportion of Wnt1.sup.+ cells ˜6.5 fold (FIG. 2 Panel C). Cells seeded without prior filtration and cultured in the same conditions also formed spheroids with a ˜57.8 fold enrichment of Wnt1.sup.+ cells, confirming that the majority of NC-derived cells originate from NFBs”). Moreover, the Specification teaches that “NC-derived Schwann cells are able to give rise to SAT-NSCs” (para 0102, Example 1), thereby indicating that neuronal stem cells (NSCs) can be isolated from Schwann cells rather than the nerve fiber bundles themselves.
Quantity of Experimentation
A person with ordinary skill in the art would not reasonably expect that culturing adipose-tissue derived nerve-fiber bundles would result in human neuronal progenitor cells. The Specification only teaches that Schwann cells within NFBs of subcutaneous adipose tissue possess neurogenic potential, rather than demonstrating that culturing nerve fibers alone without Schwann Cells will result in neuronal progenitor cells. A person with ordinary skill in the art would have to perform undue, substantial experimentation to fully encompass the scope of the claims.
Conclusion
In light of the unpredictability surrounding the claimed subject matter in the art and lack of adequate guidance in the Specification, one wishing to practice the presently claimed invention would be unable to do so without engaging in undue The claims are broadly directed to methods using any adipose tissue comprising NFBs, regardless of the presence of NC-derived Schwann cells. The prior art (Nagase et al., Lee et al.) does not teach that culturing NFBs alone, without Schwann cells, will yield neuronal progenitor cells. The specification provides insufficient guidance and only working examples for the Schwann cell-associated pathway. Therefore, a skilled artisan would need to perform undue experimentation to practice the claimed invention across its full scope. The claims are not commensurate in scope with the enabled embodiments and should be limited to methods involving NFBs containing NC-derived Schwann cells.
.
Claim Interpretation
For the purposes of examination, claim 1 is interpreted in light of the Specification, as being directed to the production of adipose-derived neuron progenitor cells from Schwann cells, located within nerve fiber bundles that were dissociated from subcutaneous adipose tissue.
Claim Rejections - 35 USC § 101
35 U.S.C. 101 reads as follows:
Whoever invents or discovers any new and useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof, may obtain a patent therefor, subject to the conditions and requirements of this title.
Claims 1-14, 16-18 and 20-21 are rejected under 35 U.S.C. 101 because the claimed invention is directed to a product of nature without significantly more. The claims recite the nature-based product drawn to adipose-derived neuronal progenitor cell. MPEP 2106 sets forth the multistep process for determining subject matter eligibility.
In accordance with the MPEP § 2016, claimed found to recite a statutory subject matter (e.g., compositions of matter) (Step 1: YES) are the analyzed to determine if the claims recite any steps that equate to an abstract idea, law of nature, or natural phenomenon. Claims 1-14, 16-18 and 20-21 are drawn a method of producing neuronal progenitor cells from adipose tissue and a product of said method, that being the human neuronal progenitor cell. Specifically, the claims recite naturally occurring cell populations isolated from adipose tissues, as discussed in Ikeya et al. (Published: 2019. CA3110026A1. Cited in IDS filed 5/29/2026). Ikeya teaches that neural precursors are cells that originated from neural crest cells (NCCs) found within the neuroectoderm (para 0002). As such, the claims and art merely recite the isolation and maintenance of such naturally occurring cell populations outside of the body, and the process of culturing does not impart any properties to the product of human neuronal progenitor cells by the claimed steps. The claimed isolated human neuronal progenitor cells have the same structure as the naturally occurring neuronal progenitor cells. They have merely been isolated and placed into culture.
MPEP 2106.04(c) states that if a claim includes a nature-based product that does not exhibit markedly different characteristics from its naturally occurring counterpart in its natural state, then the claim recites a “product of nature” exception, and requires further analysis in Step 2A Prong Two to determine whether the claim as a whole integrates the exception into practical application (Step 2A, Prong One: YES)
The judicial exception is not integrated into a practical application because they do not recite any elements in addition to the recited judicial exception. MPEP 2106.04(d), subsection III states that a judicial exception alone is not eligible subject matter; therefore, if there are no additional claim elements besides the judicial, or if the additional claim elements merely recite another judicial exception, that is insufficient to integrate the judicial exception into a practical application. The added limitations of neural progenitor cells expressing PLP1, P75, SOX10 and/or P75 and CD49f, merely identify and characterize the claimed cells based on naturally occurring characteristics rather than impose any additional steps that integrate the judicial exception.
As such, claims 1-14, 16-18 and 20-21 are directed to a natural product with no additional elements to demonstrate the claims as a whole integrate the exception into practical application. The claimed neuronal progenitor cells are isolated from adipose tissue but do not exhibit markedly different characteristics from naturally occurring neuronal progenitor cells. The process steps recited are routine and do not impart new properties. Applicant may wish to provide evidence of functional differences or amend claims to include additional steps (Step 2A, Prong 2: NO).
The claims do not include additional elements that are sufficient to amount to significantly more than the judicial exception. MPEP 2106.05, subsection I states that additional elements in the claims must be evaluated to determine whether they amount to an inventive concept, which requires considering them both individually and in combination to ensure that they amount to significantly more than the judicial exception itself. Any additional steps, such as isolating the cell populations from adipose tissue, dissociating the tissue to obtain nerve fiber bundles, are no more than routine in the art and do not transform the nature of the claim to be something distinct from their naturally occurring counterpart. Hence, because claims 1-14, 16-18 and 20-21 do not contain additional elements to demonstrate the claims as a whole integrate the exception into a practical application, the claim simultaneously does not recite significantly more than the exception itself and there is no meaningful limitation in the claim that transforms the exception into a patent- eligible application. Therefore, claims 1-14, 16-18 and 20-21 are directed to a judicial exception without significantly more and does not have an eligible subject matter under 35 U.S.C. § 101 (Step 2B: NO).
Claim Rejections - 35 USC § 103
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.
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.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claim(s) 1-3, 6, 8, 9-11 is/are rejected under 35 U.S.C. 103 as being unpatentable over Nagase et al. (Published: 2008. WO2008150001A1. Cited in IDS filed 04/03/2025. Machine Translation provided by Examiner) and Lee et al. (Published 2021. International journal of molecular sciences, 22(8), 4141.) and Chen et al. (Published: 2017. American journal of translational research, 9(5), 2579–2594.)
Regarding claim 1, Nagase teaches producing human neuronal progenitor cells from adipose tissue (para 0020, " The cells derived from adipose tissue are cells derived from fat under the skin, in the groin, or in the abdominal cavity, as described in the above items...The cells derived from adipose tissue are cells possessing pluripotency, according to the composition described above."; para 0022, "This aggregation can be used for regenerative therapy. Furthermore, the present invention can prepare cells with the function of neural stem cells by recovering precursor cells from fat and applying the conditions for aggregation of these precursor cells.”). Nagase teaches dissociating fat (adipose) tissue via collagenase treatments (para 0020), The method described above comprises a step in which fat tissue or aspirated fat waste liquid is collected before step A), collagenase treatment is performed, then centrifuged and collected as pellets to obtain cells derived from adipose tissue, (Claim 5, "The adipose tissue-derived cells are either stromal-vascular-fraction cells for which adipose tissue is harvested, collagenase treatment, centrifuged and collected as pellets, or adipose-derived stem cells derived from waste fluid, according to claim 1.") and culturing the cells to obtain adipose derived-stem cells (para 0020, step 25 and 26, "After step A), A') the step of adhering cells derived from adipose tissue, according to the above item. (26) The method according to the above item, wherein following step A') the adipose tissue-derived cells are adhered to A'), and thereafter subsequently passaging once or more to obtain the fat tissue-derived cells as adipose-derived stem cells (ASC).").
However, Nagase fails to teach collecting NFBs from adipose tissue.
Chen teaches that adipose tissue contained nerve fibers and that Schwann cells can be isolated from adipose tissue since nerve fibers are found within adipose tissue (pp. 1-2 “In addition, many studies have also demonstrated that adipose tissue contains various cell components [19] and auto nomic nerves [20-23] Since nerve fibers are found in adipose tissue, it is possible to collect SCs from adipose tissue.”). Moreover, Chen teaches dissolving adipose tissue to obtain Schwann-like cells (“Tissues were placed in a 50 ml conical centrifuge tube containing an enzymatic solution (50 μl per segment) prepared by dissolving Collagenase NB4 (Serva, Germany) and DispaseII (Serva) into DMEM at a concentration of 0.2% (0.27 U/ml).”).
The combined teachings of Nagase and Chen do not teach culturing nerve fiber cells from adipose tissue.
Lee teaches a method of isolating peripheral-nerve stem cells from nerve fragments (Abstract, “Here, we isolated novel peripheral nerve-derived stem cells (PNSCs) from adult peripheral nerves with similar characteristics to neural-crest stem cells.”). Further, Lee teaches that neural crest stem cells migrate along nerve fibers (pp. para 2, “The neural crest stem cells (NCSCs) have been shown to migrate along nerve fibers for wide distribution, where they give rise to a variety of neural crest (NC)-lineage cells ranging from neurons and glia of the peripheral nervous system to non-neural cells, including melanocytes, endocrine cells, endoneuria (from the endoneurium) fibroblasts, pericytes, chondrocytes, osteoblasts, and MSCs”).
It would have been obvious to combine the progenitor cell culture method of Nagase with an additional step of further collecting and culturing nerve fiber bundles to obtain neuronal progenitor cells, as Chen teaches that nerve fibers are found within adipose tissue, and Lee teaches a method of isolating stem cells (e.g., neural crest stem cells) from nerve fibers as Lee teaches that neuronal differentiation potential of nerve fibers comprising neural crest stem cells (NCSCs) . A skilled artisan would have recognized, in view of Chen and Lee, that adipose tissue contains nerve fibers that are associated with stem cells, and that such neural crest stem cells (e.g, neuronal progenitor cells) can be isolated to obtain neuronal progenitor cells with a reasonable expectation of success.
Regarding claim 2, the combined teachings of Lee, Nagase and Chen render obvious the claimed method of claim 1. Moreover, Nagase teaches culturing until spheroids are formed (para 0012, "Traditionally, the suspension culture method on neurosphere [spheroid] media was recognized as a method for efficiently isolating and proliferating so-called neural stem cells, and the principle was that it was a cloning cell growth mass from a single cell."; para 0201, "Against this backdrop, the inventors floated and cultured high-density adipose-derived cells on specially coated non-adhesive plates in neurosphere medium, and found that sphere-like cell masses obtained in 1-3 days were positive for neural stem cell markers such as nestin and Musashi-1.").
Regarding claim 3, the combined teachings of Lee, Nagase and Chen render obvious the claimed method of claim 1. Moreover, Nagase teaches counter-filtration (para 0178, " The supernatant was aspirated, and precipitate cells were collected to a total volume of 15~20ml. If a large amount of matrix components was present, the matrix components were filtered and removed using a 100μm filter. Subsequently, centrifugation was performed as needed.").
Regarding claim 6, the combined teachings of Lee, Nagase and Chen render obvious the claimed method of claim 1 and 2. Moreover, Nagase teaches expanding the spheroids (para 0062, "dissociating the formed neurosphere and seeding the dissociated cells into a new medium, and a method that includes the step of verifying whether these dissociated cells form a neurosphere again. In this method, when the dissociated cells reform a neurosphere, they are identified as neural stem cells.").
Regarding claim 8, the combined teachings of Lee, Nagase and Chen render obvious the claimed method of claim 1. Moreover, Nagase teaches enzymatic digestion via collagenase treatment (Claim 5, "The adipose tissue-derived cells are either stromal-vascular-fraction cells for which adipose tissue is harvested, collagenase treatment, centrifuged and collected as pellets, or adipose-derived stem cells derived from waste fluid, according to claim 1.") and filtering the adipose tissue (para 0143, “The above sample is suction-filtered using a mesh with a diameter of 100 μm; and (8) the obtained filter can be separated by centrifugation at 1200 g for 5 minutes.”).
Regarding claim 9, the combined teachings of Lee, Nagase and Chen render obvious the claimed method of claim 1 and 8. Moreover, Nagase teaches that the adipose tissue was harvested using collagenase (claim 16, "The adipose tissue-derived cells are either stromal-vascular-fraction cells for which adipose tissue is harvested, collagenase treatment, centrifuged and collected as pellets, or adipose-derived stem cells derived from waste fluid, according to claim 1.").
Regarding claim 10, the combined teachings of Nagase, Lee, Chen and Ikeya render obvious the claimed method of claim 1, 8 and 9. Moreover, Chen teaches that the adipose tissue was harvested using dispase and collagenase (para 0040, “For example, a method of cutting an intestinal tract tissue into 1 mm square, carrying out enzymatic treatment (Dispase and collagenase type XI, 37 C, 90 min), and then culturing the obtained cells in a medium containing EGF and FGF to separate ENPs as aggregated cells is known as a method for separating ENPs from a living body.”)
Regarding claim 11, the combined teachings of Lee, Nagase and Chen render obvious the claimed method of claim 1. Moreover, Nagase teaches subcutaneous adipose tissue (para 0025, "Figure 2 shows adipoclusters derived from subcutaneous fat and intraperitoneal fat in Nestin EGFP transgenic rats. Although the test was done one day later using method (1), it is already positive for EGFP signaling and considered to have increased nestin expression."; para 0083, "this specification, "adipose tissue" refers to loose connective tissue, particularly those with a high number of adipocytes, each fat cell surrounded by lattice fibers, and capillaries densely distributed between the cells. It is also called the fat body. Typically, it refers to subcutaneous fat tissue").
***
Claim(s) 1, 4 and 5 is/are rejected under 35 U.S.C. 103 as being unpatentable over Nagase et al. (Published: 2008. WO2008150001A1. Cited in IDS filed 04/03/2025. Machine Translation provided by Examiner),Lee et al. (Published 2021. International journal of molecular sciences, 22(8), 4141.) and Chen et al. (Published: 2017. American journal of translational research, 9(5), 2579–2594.), in further view of Brown et al. (Published: 2013. Journal of neuroscience methods, 209(2), 344–350).
With regard to claim 1, the combined teachings of Nagase, Lee and Chen render obvious the claimed methodology, as iterated above in the 103 rejection the content of which is incorporated in claim 1. Moreover, Chen teaches that nerve fibers were visualized using Hematoxylin and Eosin (H&E) staining (pp. 2584 col 1-col2, “To investigate the existence of nerve fibers and SCs in adipose tissue, we performed H&E stain 2584 ing, IF staining and TEM of inguinal adipose tis sue. H&E staining showed that some structures looked similar to nerve fibers in adipose tissue”).
Regarding claims 4 and 5, the combined teachings fail to disclose that the nerve NFBs are visualized with a dye, specifically FluoroMyelin red.
Brown teaches that fluoromyelin provides live labelling of myelin sheaths or axons within nerve fibers (pp. 345 col 1, “ We show that FluoroMyelin™ Red provides bright, photostable and selective fluorescent labeling of myelin in live myelinating cultures with no apparent toxicity and no apparent adverse effects on the myelin sheaths or the axons that they invest.”).
Therefore, it would have been obvious to substitute Brown’s fluoromyelin for Chen’s identification of nerve fibers through H&E staining as fluoromyelin provides fluorescent labelling of live nerve fibers, providing more specificity or selectivity for those cell populations. There would have been reasonable expectations of success in combining these teachings as one of ordinary skill in the art would recognize to combine known elements in the art to give predictable results.
***
Claim(s) 1, 7, 13, 14, 16-18 and 20-21 is/are rejected under 35 U.S.C. 103 as being unpatentable over Nagase et al. (Published: 2008. WO2008150001A1. Cited in IDS filed 04/03/2025. Machine Translation provided by Examiner),Lee et al. (Published 2021. International journal of molecular sciences, 22(8), 4141.) and Chen et al. (Published: 2017. American journal of translational research, 9(5), 2579–2594.), in further view of Ikeya et al. (Published: 2020. CA3110026. Provided by Examiner).
With regard to claim 1, the combined teachings of Nagase, Lee and Chen render obvious the claimed methodology, as iterated above in the 103 rejection the content of which is incorporated in claim 1. Moreover, Lee teaches a cell culture medium containing fetal bovine serum (pp. 2580, col 1).
However, the combined teachings fail to disclose that the NFBs are cultured on a fibronectin-coated substrate, required by claim 7.
Ikeya teaches a method of producing neuronal precursor cells , further comprising a surface-treated culture container coated with fibronectin to improve adhesiveness to cells (para 0074, “The container for use in adherent culture may be surface-treated in order to improve adhesiveness to cells (hydrophilicity), or coated with a substrate for cell adhesion such as collagen, gelatin, poly-L-lysine, poly-D-lysine, laminin, fibronectin, Matrigel, or vitronectin.”)
It would have been obvious to modify the method of producing neuronal progenitor cells of Nagase, to further comprise culturing the NFBs on a fibronectin coated culture substrate as taught by Ikeya, as a fibronectin-coated culture container would improve the adhesiveness of culturing cells to a culture container. There would have been reasonable expectations of success in combining these teachings as one of ordinary skill in the art would recognize to combine known elements in the art to give predictable results.
Regarding claim 13, 14, and 16 the combined teachings of Nagase, Lee and Chen render obvious the method of claim 1. Moreover, Nagase teaches that the neural crest cell markers SOX10 and p75NTR (para 0068-0069).
Further, Ikeya teaches that neural precursor express PLP-1 (para 0018, “ ENPs have differentiation capacity into PHOX2B-positive and Date Recue/Date Received 2021-02-18 SOX10-negative enteric nerve cells, and S100-positive, PLP1-positive and SOX10-positive glial cells.”).
Therefore, it would have been obvious for one of ordinary skill in the art to identify neural crest cells by the combinations of at least 3 markers SOX10, p75NTR and PLP1 with a reasonable expectation of success as these markers characterize neuronal progenitor cells.
Note, in regards to claim 16, the Nagase and the cited prior art is silent on the expression of ZIC1. Thus, the combined teachings of Lee, Chen, Nagase and Ikeya do not teach characterization of neuronal progenitor cells using ZIC1, absent factual evidence to the contrary.
Regarding claim 17 the combined teachings of Nagase, Lee and Chen and Ikeya render obvious the method of claim 1 and product of claim 14. Moreover, Ikeya teaches administering a cell medicament comprising the neural precursor cells to the stomach or colon, (para 0099, “Examples of the method for administering the cell medicament include intraperitoneal injection, subcutaneous injection, injection into the lymph node, intravenous injection, intrathoracic injection, direct injection to a local gastrointestinal organ (for example, the esophagus, the stomach, the duodenum, the small intestine, the jejunum, the ileum, the colon, and the rectum) by opening the abdomen, and administration into the rectal cavity.”; para 0146, “The enteric neural precursors were confirmed to be engrafted in the mouse intestinal tract”).
Regarding claim 18 the combined teachings of Lee, Chen, Nagase and Ikeya render obvious the claims 1, 14 and 17. Moreover, Ikeya teaches a method for treating Hirschsprung disease by transplanting enteric precursor cells (para 0003, “In recent years, a fundamental approach aimed at treating Hirschsprung disease by autotransplanting ENPs that have collected Date”, para 0095, “ENPs obtained by the production method or the expansion culture method according to the present invention may be applied to a cell medicament for the prevention or treatment of a disease caused by deficiency or abnormality in enteric nerve cells. Examples of such a disease include Hirschsprung disease…”).
Regarding claim 20 and 21, the combined teachings of Lee, Chen, Nagase and Ikeya render obvious the claims 1 and 14. Moreover, Lee teaches contacting spinal cords (central-nervous system) with peripheral-nerve spheroids to treat spinal cord injury (Abstract, “In an animal spinal cord injury (SCI) model, these PNSC spheroids induced functional recovery and neuronal regeneration.”).
***
Claim(s) 1 and 12 is/are rejected under 35 U.S.C. 103 as being unpatentable over Nagase et al. (Published: 2008. WO2008150001A1. Cited in IDS filed 04/03/2025. Machine Translation provided by Examiner),Lee et al. (Published 2021. International journal of molecular sciences, 22(8), 4141.) and Chen et al. (Published: 2017. American journal of translational research, 9(5), 2579–2594.) as applied to claim 1 above and further in view of Blaszkiewicz et al. (Published: 2019. Biology, 8(1), 10.).
With regard to claim 1, the combined teachings of Nagase, Lee and Chen render obvious the claimed methodology, as iterated above in the 103 rejection the content of which is incorporated in claim 1. In particular, Nagase teaches subcutaneous adipose tissue (para 0025), However, the combined teachings do not disclose isolating progenitor cells from visceral adipose tissue.
Blaszkiewicz teaches that nerve fibers are found within visceral adipose tissue (pp. 8, para 5, “This study found significant intra-adipose variation in TH+ fibers between subcutaneous and visceral depots”).
It would have been obvious to substitute Blaszkiewicz’s visceral adipose tissue for Nagase’s subcutaneous adipose tissue. Therefore, these compositions are functional equivalents in the art, and substituting one for the other would have been obvious at the time of the invention. “When a patent ‘simply arranges old elements with each performing the same function it had been known to perform’ and yields no more than one would expect from such an arrangement, the combination is obvious.” See KSR International Co. v. Teleflex Inc., 82 USPQ2d 1385 (U.S. 2007) at 1395-1396, quoting Sakraida v. AG Pro, Inc., 425 U.S. 273 (1976) and In re Fout, 675 F.2d 297, 301 (CCPA 1982) (“Express suggestion to substitute one equivalent for another need not be present to render such substitution obvious”). There would have been reasonable expectations of success as using obtaining adipose tissue from different sources was well known in the art before the effective filing date of the claimed invention.
Conclusion
No claims allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to Katriel B Kasayan whose telephone number is (571)272-1402. The examiner can normally be reached 10-4p.
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If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Maria G Leavitt can be reached at (571) 272-1085. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/KATRIEL BARCELLANO KASAYAN/Examiner, Art Unit 1634
/MARIA G LEAVITT/Supervisory Patent Examiner, Art Unit 1634
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