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
Acknowledgement is hereby made of receipt and entry of the communication filed on July 24, 2026. Claims 17, 24 and 27-28 are pending and currently examined.
Claim Rejections - 35 USC § 112 (d)
The following is a quotation of 35 U.S.C. 112(d):
(d) REFERENCE IN DEPENDENT FORMS.—Subject to subsection (e), a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
The following is a quotation of pre-AIA 35 U.S.C. 112, fourth paragraph:
Subject to the following paragraph [i.e., the fifth paragraph of pre-AIA 35 U.S.C. 112], a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
(New rejection) Claim 17 is rejected under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends, or for failing to include all the limitations of the claim upon which it depends. Claim 17 is improperly dependent on claim 24 after claim 17. 35 U.S.C. § 112(d) states that “a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed.
Applicant may cancel the claim(s), amend the claim(s) to place the claim(s) in proper dependent form, rewrite the claim(s) in independent form, or present a sufficient showing that the dependent claim(s) complies with the statutory requirements.
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.
(Previous rejection- withdrawn) Claims 1, 7, 12-14 and 16
This rejection is withdrawn in view of the amendment filed on July 24, 2026.
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.
(Previous rejection- withdrawn) Claim 18 is rejected under 35 U.S.C. 103 as being unpatentable over Allbritton et al. (US 11193110 B2, patented on Dec. 7, 2021 and PCT filed on Jan. 29, 2016) as applied to claims 1, 7, 12-14 and 16 above.
This rejection is withdrawn in view of the amendment filed on July 24, 2026.
(Previous rejection-withdrawn) Claims 19 and 27 are rejected under 35 U.S.C. 103 as being unpatentable over Allbritton et al. (US 11193110 B2, patented on Dec. 7, 2021 and PCT filed on Jan. 29, 2016) in view of Sato et al. (WO 2010/090513 A2, published on Aug. 12, 2020) as applied to claims 1, 7, 12-14, 16-18 and 24 above and further in view of Chang et al. (Proc Natl Acad Sci U S A. 2004 Jun 8;101(23):8733-8.).
This rejection is withdrawn in view of the amendment filed on July 24, 2026.
(New Rejection-necessitated by amendment) Claims 17 and 24 are rejected under 35 U.S.C. 103 as being unpatentable over Allbritton et al. (US 11193110 B2, patented on Dec. 7, 2021 and PCT filed on Jan. 29, 2016) and in view of Sato et al. (WO 2010/090513 A2, published on Aug. 12, 2020), and Karst et al. (WO 2015/175526 A2, published on Nov. 19, 2015).
Regarding the base claim 24, Allbritton et al. teaches the claim 24 (1) by stating that to obtain the 3D organoid from human intestine and cultured in the 3D organoid model to expand the cells, including stem cells, progenitor cells, differentiated cells and human colonic epithelial cells (See Example 5; Figure 10 below), where the isolated crypts, isolated stem cells or colonic epithelial cells are within a 3D extracellular matrix (ECM) (See column 1, lines 59-67; Column 4, Fig. 8), and where a cell culture medium comprising a Wnt agonist (R-spondin, an epidermal growth factor (EGF), a transforming growth factor-(TGF- β) such as A83-01 and BMP inhibitor such as Noggin (See column 13, lines 1-12; column 20, lines 9-32; column 9, lines 13-41).
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As for the claim 24 (2), the limitation for preparing the 2D organoid from dispersing the 3D organoid to a single cell is taught by Allbritton that the cell fragments grew into 3D organoids at day 2 and continued to expand into large organoids at day 6. In addition, the cells in the 3D organoids so prepared could be switched back to grow as a 2D monolayer by releasing organoids from Matrigel by repetitive mechanical pipetting, and plating the organoids on the collagen scaffold (FIG. 4C). The organoids spread on the scaffold at day 1 and formed a 2D monolayer at day 6 (FIG. 4C) (See column 14), where it is reasonable to consider that the repetitive mechanical pipetting can disperse the 3D organoid to prepare a single cell as claimed. Allbritton also teaches that a 2D colonic epithelial monolayer was generated by culturing isolated crypts or stem cells on the surface of the planar, 2D scaffolds, while the biophysical microenvironment was provided by the collagen scaffold which mimics the lamina propria in terms of permeability, stiffness, and ECM components (See column 13, lines 1-9 and Figure 1 A below). Here the single cell is shown in Figure 1A below and Figure 1A also shows that there is no lumen in the 2D organoid. Allbritton discloses that self-renewing gastrointestinal epithelial cell monolayer extending onto said well side walls and floors, with the well top openings remaining open, to form open lumens lined with cells (See column 26, claim 10).
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As for the limitation that “…epithelial cells contain differentiated trophoblastic cells and goblet cells” in claim 24 (2), Allbritton teaches the differentiated goblet cells (See Figure 5 below and column 4, lines 1-3).
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For the trophoblastic cells, Allbritton et al. does not explicitly point out the “trophoblastic cells”. However, Allbritton et al. teaches using the same method as claimed to acquire 2D culture under the same culture medium, it is reasonable to expect that the intestinal epithelial cell 2D organoids disclosed in Allbritton are indistinguishable from that of the instant invention. The Office does not have the facilities and resources to provide the factual evidence needed in order to establish that the intestinal epithelial cell 2D organoid derived from human intestinal epithelial stem cells, human intestinal epithelial cell or a tissue containing at least one of these disclosed in Allbritton does not contain the differentiated trophoblastic cells. In the absence of evidence to the contrary, the burden is on the applicant to prove that the claimed invention is different from those taught by the prior art and to establish patentable differences. See In re Best 562F.2d 1252, 195 USPQ 430 (CCPA 1977) and Ex parte Gray 10 USPQ 2d 1922 (PTO Bd. Pat. App. & Int. 1989).
For the culture medium in claim 24-(2), Allbritton teaches a 2D monolayer culture medium containing i) Wnt agonist (Wnt-3A, R-spongin), ii) EGF, iii) BMP inhibitor/noggin, iv) TGF-β inhibitor/A83-01 and v) a Rock inhibitor/ Y27632 (See e.g., column 20, Example 5).
Accordingly, Allbritton teaches the claim 24-(2).
As for the method of producing a 2D organoid culture for use in infection and replication of human norovirus as claimed, Allbritton et al. teaches that their invention cab be used for screening studies of drugs, biologics, toxins, mutagens, dietary compounds, pathogens, viruses, microbiota, etc. (See column 11). However, it does not specifically point out culturing the human norovirus.
Nevertheless, Sato et al. teaches using the crypt-villus, gastric or pancreatic organoids to culture a pathogen such as a norovirus which presently lacks a suitable tissue culture or animal model (See page 22, lines 26-28), which teaches that the 2D organoid can be used for growing norovirus as claimed in the instant claim 24 (2).
Sato et al. also teaches the “single cell” claimed in the instant claim 24 (2) by stating that single cell suspension comprising the epithelial stem cells is mechanically generated from the isolated crypts (See e.g., page 4, lines 14-24) and a scaffold provides a two-dimensional or three-dimensional network (See page 5, lines 6-15). Sato et al. also teaches seeding 100 single cells/well and counted the number of organoids 14 days after seeding (See e.g., page 48, lines 13-21). Here it indicates a process of dispersing the 3D organoid to seed a single cell on the plate for growing the 2D monolayer culture as described as “gastric cultures have been grown in monolayers” (See page 52, lines 15-28).
In addition, Karst teaches that the norovirus can be replicated in vitro in a cell culture such as intestinal epithelial cells, where continuous culture of norovirus comprises a) contacting a norovirus to cells in a transwell in vitro cell culture system comprising a transwell culture vessel comprising intestinal epithelial cells grown on a transwell membrane (See page 13).Here the intestinal epithelial cells grown on a Transwell membrane can be a 2D organoid. Here the norovirus is human norovirus (page 2, lines 27-29). Karst’s disclosure is providing an in vitro system using 2D culture to grow and replicate the norovirus in a and aimed at making norovirus easier to detect, study, and grow continuously.
It would have been obvious to one of ordinary skill in the art at the time of the invention to introduce the teaching of Sato and Karst into Allbritton’s invention to culture the norovirus using the 2D organoid. One would have been motivated to do so because Sato teaches using a single cell for starting the 2D culture thus generating a uniform and long-last organoid. It is obvious a uniform organoid and long-last culture would be benefitting the norovirus culturing. Karst teaches using an intestinal epithelial cell in a transwell to grow norovirus in an in vitro cell culture system efficiently (See page 28, lines 19-33). Therefore, there would have been a reasonable expectation of success to develop a method of producing a 2D organoid culture for norovirus infection and replication.
Also, the above description teaches claim 17 for producing norovirus using a 2D organoid culture.
Thus, the invention as a whole was clearly prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention.
(New Rejection-necessitated by amendment) Claim 27 is rejected under 35 U.S.C. 103 as being unpatentable over Allbritton in view of Sato and Karst as applied to claims 17 and 24 above and further in view of Chang et al. (Proc Natl Acad Sci U S A. 2004 Jun 8;101(23):8733-8.).
Claim 27 requires that the cell medium further contains animal-derived bile.
Relevance of Albritton in view of Sato and Karst is set forth above. However, it is silent on the use of animal-derived bile.
Chang et al. teaches that a porcine enteric calicivirus (PEC), strain Cowden in the family Caliciviridae (genus Sapovirus), can be propagated in a continuous cell line, LLC-PK cells, but only in the presence of an intestinal content fluid filtrate from gnotobiotic pigs. This cell culture system is presently the only in vitro model among caliciviruses that cause gastrointestinal disease, including members of the genera Sapovirus and Norovirus. They report here the identification of bile acids as active factors in intestinal content fluid essential for porcine enteric calicivirus replication in association with down-regulation of signal transducer and activator of transcription 1 and proposes a mechanism for enteric calicivirus growth mediated by bile acids, ubiquitous molecules present in the intestine at the site of the virus replication, that is associated with inhibition of innate immunity (See Abstract; page 8736, right column, paragraph 1). Chang et al. also discloses that the effects of various bile acids derived from animal on PEC growth in cell culture, where the bile acids include Porcine bile extract and bovine bile extract (See Fig. 2, page 8735).
Accordingly, Chang’s study indicates that the bile acids derived from porcine is an essential component for enteric virus (including norovirus) replication and growing.
It would have been prima facie obvious for one of ordinary skill in the art at the time of invention to introduce porcine derived bile acids into the inventions of Allbritton Sato Karst when culturing and growing norovirus viruses. One would have been motivated to do so because Chang teaches that bile is essential for norovirus virus replication and can be applied to the development of cell culture systems for nonculturable viruses (See page 8738, last paragraph). There is a reasonable expectation of success to add bile acids into the culture medium to facilitate production of human norovirus as claimed in the instant application.
(New Rejection-necessitated by amendment) Claim 28 is rejected under 35 U.S.C. 103 as being unpatentable over Allbritton in view of Sato and Karst as applied to claims 17 and 24 above, and further evidenced by Clevers et al. ( US 2014/0243227 Al, published on Aug. 28, 2014).
Regarding claim 28, it requires the extracellular matrix is basement membrane preparation.
Allbritton et al. teaches using the extracellular matrix for support of the organoids and the colonic epithelial cells cultured under ENR-WB for 5 days on Matrigel/collagen scaffold (See e.g., Fig. 8). Here Clevers’ invention evidence that Matrigel is a solubilized basement membrane preparation extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma (See [0129]).
Responses to Applicant’s Remarks
Applicant’s arguments filed on July 24, 2026, have been received and fully considered.
Because applicant canceled claims 1, 7, 12-14 and 18-19, the rejection under 35 U.S.C. § 102(a)(1) for claims 1, 7 and 12-14 is withdrawn, and the rejections under 35 U.S.C. § 103 for claims 18-19 are withdrawn.
Applicants’ argument regarding claims 17 and 24 are not found persusasive.as follows:
(1). Applicant argued that Allbritton and Sato do not teach or suggest either step (1) or step (2) of amended independent claim 24 (See pages 6-8).
The argument is not persuasive.
(a) Based on the description above, Allbritton teaches the amended claims 24 (1) and (2), for example, Allbritton teaches stem cells could be obtained from biopsy of a patient with digestive epithelial damage (See bringing pages of column 11-12), and Figure 10 (See above) teaches a preparation of 2D monolayers from the 3D organoid (answered Remarks, page 6).
(b) Allbritton teaches the cells in the 3D organoids so prepared could be switched back to grow as a 2D monolayer by releasing organoids from the Matrigel by repetitive mechanical pipetting and plating the organoids on the collagen scaffold (See column 14, lines 60-67), which implies the “dissociating it into single cells” of the 3D organoid can occur (answered Remarks, page 6). Figures 10 and 1 above also show the monolayer culturing of the 3D organoid.
(c) Applicant argued the new culture medium and the addition of Rock inhibitor (See Remarks, page 6). The argument is not persuasive.
Allbritton teaches that media were changed daily (See column 17, lines 15-20). After 3D organoids are mechanically dispersed, many factors are added into the medium for the transition from 3D organoid to 2D organoid, for example, adding Wnt-3A and Notch inhibitors for Goblet cell differentiation (See column 3, lines 65-67), also, when spreading 2D monolayer possessing an exposed luminal surface, the Rock inhibitor is added (See column 20, lines 25).
Accordingly, Allbritton teaches generating the 2D organoid from generating the 3D organoid and Rock inhibitor is added for conversion of the 2D monolayer. In addition, Sato also teaches that a further embodiment of a method according to the invention comprises a culture medium comprising a Rock (Rho-kinase) inhibitor. The addition of a Rock inhibitor was found to prevent anoikis, especially when culturing single stem cells (See Sato, page 11, lines 23-25).
(2) Applicant argued that the Allbritton does not teach changing the culture medium when transitioning from 3D to 2D (See Remarks, page 7).
The argument is not persuasive.
Allbritton’s invention does not indicate that a culture medium was kept constant. Actually, Allbritton teaches the media being changed daily and different reagents such as Wnt3a, notching inhibitor and Rock inhibitor are added for the conversion.
Additionally, applicant’s arguments on “changing the culture medium at the 2D stage” and “changing the culture medium in the conversion of a 3D organoid to a 2D organoid” are not required in the instant claims.
Applicant argued that Allbritton also does not teach a production method in which, when generating 3D organoids and converting them into 2D monolayer, a Rock inhibitor is added to the medium upon dissociation into single cells (See Remarks, page 8).
The argument is not persuasive.
Allbritton’s example 5 teaches adding the Rock inhibitor in the converted 2D monolayers. Although there is mentioned “single cells” in example 5, Allbritton teaches using the mechanical pipetting tool to prepare the 2D monolayers from the 3D organoid (See column 14, lines 64). It is obvious that medium culture of the organoid includes the single cells (See Figure 1 above). In addition, Sato teaches that “For the single cell culture, a total of 100 sorted EGFPhi cells /well were collected in gastric culture medium and embedded in Matrigel (BO Bioscience). After polymerization of Matrigel, gastric culture media was overlaid. For the first 2 days after seeding, the media was also supplemented with 10μM ROCK inhibitor Y-27632 (Sigma Aldrich), to avoid anoikis (See Sato, page 71, lines 24-29).
(3) Applicant argued that the claimed method provides significant norovirus replication (See Remarks, page 8).
The argument is not persuasive.
The instant claims do not cite a limitation on the “significant norovirus replication”. Therefore, the arguments such as “A 2D organoid was formed and infected with norovirus by cutting the 3D organoid using TrypLE Express to make a single cell” (See Remarks, page 9), and “robust replication of norovirus in a 2D human epithelial model” (See Remarks, page 10) are not part of the claims.
Applicant argued the reference of Chang and argued that a person of ordinary skill in the art would have no suggestion or motivation with a reasonable expectation of success that the same cascade would operate or even be present in a norovirus physiological target system (i.e. human small-intestinal epithelium) (See Remarks, page 11).
The argument is not persuasive.
Chang is used for addressing a specific limitation for the cell culture medium containing animal-derived bile and teaches that bile acids are essential for porcine enteric calicivirus replication. It is proper to be used as a support reference to Allbritton in view of Sato.
Applicant argued against the reference of Clevers by allegedly saying claim 28 is not obvious over Allbritton in view of Sato and further in view of Clevers.
The argument is not persuasive.
Based on the amendment, Allbrittton teaches monolayer-culturing the 3D organoid on the extracellular matrix Matrigel (See column 8, lines 20-30), where Clevers is applied in the current office action to be used as evidence to demonstrate that Matrigel is a basement membrane preparation from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells (See Clevers, [0129]).
Conclusion
No claims are allowed.
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 extension fee 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 date of this final action.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to RUIXUE WANG whose telephone number is (571)272-7960. The examiner can normally be reached Monday-Friday 8:00 am-5:00 pm, EST.
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/RUIXUE WANG/ Examiner, Art Unit 1672
/NICOLE KINSEY WHITE/ Primary Examiner, Art Unit 1672
