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 .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on Feb. 11, 2026.
Detailed Action
Acknowledgement is hereby made of receipt and entry of the communication filed on Feb. 11, 2026. Claims 1, 7, 12-14, 16-19, 24, 27 and 28 are pending and currently examined.
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.
(New rejection) Claims 1, 7, 12-14 and 16
The base claim 1 is directed to a 2D organoid culture or a method of producing a 2D organoid culture for infection and replication of human norovirus, obtained by:
(1) obtaining a 3D organoid by three-dimensionally culturing a human intestinal epithelial stem cell, a human intestinal epithelial cell, or a tissue containing at least any of those cells on an extracellular matrix; and
(2) dispersing the 3D organoid to prepare a single cell, and monolayer-culturing the single cell on an extracellular matrix-coated plate to obtain the 2D organoid having a monolayer structure adhered flatly without lumen on the plate in which epithelial cells contain differentiated trophoblastic cells and goblet cells configured as human intestinal lumen having a monolayer structure on a plate and inner part of an intestinal tract is exposed,
wherein steps (1) and (2) comprise a cell culture medium comprising i) a Wnt agonist, ii) an epidermal growth factor (EGF), iii) a bone morphogenetic protein (BMP) inhibitor and iv) a transforming growth factor- (TGF- β) inhibitor is used, and wherein the Wnt agonist is R-spondin and the BMP inhibitor is noggin.
The base claim 1 and claims 7, 12-14 and 16 are product-by-process claims, which are drawn to a 2D organoid culture. The method steps recited to obtain the 2D organoid culture are not considered when determining patentability of the product (MPEP § 2113). In In re Thorpe, the court stated, “even 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). (Emphasis added)
Allbritton et al. teaches that stem/progenitor cells make up the 2D organoid on scaffolds (See e.g., column 14, lines 14-19) and the human small intestinal epithelial cells can be cultured as 2D monolayers (See column 20, lines 5-32). Allbritton et al. also teaches that the 2D monolayer can be selectively and fully differentiated into mature enterocyte and goblet cells and the 2D monolayer on collagen scaffolds cultured under ENR-WB condition possesses stem/progenitor cells and is therefore self-renewing (See column 15, lines 4-19). Allbritton et al. discloses that the 2D organoid possesses in Vivo-Like proliferating capability, morphology and function (See Example 2) and can be used for screening a test compound or microbe for a toxicological, physiological, or carcinogenic effect (See column 12).
As for the phrase “for infection and replication of human norovirus”, it is an “intended use”. Based on MPEP 2111.02, the “intended use” does not add a patentable limitation if the structure of the device is already known in the prior art. Here Allbritton teaches the claimed 2D organoid. Nevertheless, Allbritton teaches that their invention including the 2D organoid provides an in vitro model for screening studies of drugs, biologics, toxins, mutagens, dietary compounds, pathogens, viruses, microbiota, etc. (See column 11, lines 8-16) and the microbe is an enteric bacteria or pathogen, including both benign and infectious enteric bacteria and pathogen (See column 12, lines 29-41).
Accordingly, the invention of Allbritton anticipates the claimed 2D organoid in claims 1, 7, 12-14 and 16
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.
(New rejection) 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.
Regarding claim 18, it requires a culture kit for a 2D organoid culture for infection and replication of human norovirus according to Claim 1, comprising: a cell culture medium comprising i) a Wnt agonist, ii) an epidermal growth factor (EGF), iii) a (bone morphogenetic protein (BMP) inhibitor, and iv) a transforming growth factor-n (TGF- β) inhibitor, wherein the Wnt agonist is R-spondin and the BMP inhibitor is noggin.
Based on the description above, Allbritton et al. teaches producing a 2D organoid from the 3D organoid. For example, Allbritton teaches obtaining the 2D organoid from the 3D organoid like from the Human Small Intestinal Epithelial Cells under the culturing conditions including R-spondin, noggin, EGF, BMP inhibitor and A83-01 as claimed. Although Allbritton does not explicitly specify a “kit”, the concept of packaging components into a kit is well known and routine in the art. For example, Allbritton teaches the method and needed components for producing a 2D organoid, it would have been obvious to one of ordinary skill in the art at the time the invention was made to package components into a kit. One would be motivated to do this for commercial exploitation of the invention by providing convenience for the end user. Thus, the claimed invention is obvious over Allbritton and Sato’s teachings.
(New rejection) 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).
Regarding claim 17, it is directed to a method of producing norovirus, comprising: infecting the 2D organoid culture for infection and replication of human norovirus according to Claim 1 with human norovirus and culturing the infected 2D organoid to replicate the norovirus. Based on the description above, Allbritton teaches a 2D organoid obtained from a 3D organoid as claimed and further teaches that their invention provides an in vitro model for screening studies of drugs, biologics, toxins, mutagens, dietary compounds, pathogens, viruses, microbiota, etc. (See column 11, lines 8-16) and the microbe is an enteric bacteria or pathogen, including both benign and infectious enteric bacteria and pathogen (See column 12, lines 29-41).
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).
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 into Allbritton’s invention to culture the norovirus using the 2D organoid. One would have been motivated to set up an experiment to culture the human norovirus in 2D organoid of Allbritton based on Sato’s teaching because in vitro amplification of human norovirus is a challenge due to the lack of cell culture or animal infection models and it is most important for the dearth of understanding of human norovirus. At the same time, Allbritton teaches obtaining the 2D organoid from the crypts isolated from human intestine. There would have been a reasonable expectation of success to culture the human norovirus as claimed based on the teachings of Albritton and Sato.
Regarding the base claim 24, it is directed to a method of producing a 2D organoid culture for infection and replication of human norovirus, obtained by:
(1) obtaining a 3D organoid by three-dimensionally culturing a human intestinal epithelial stem cell, a human intestinal epithelial cell, or a tissue containing at least any of those cells on an extracellular matrix; and
(2) dispersing the 3D organoid to prepare a single cell, and monolayer-culturing the single cell on an extracellular matrix-coated plate to obtain the 2D organoid having a monolayer structure adhered flatly without lumen on the plate in which epithelial cells contain differentiated trophoblastic cells and goblet cells configured as human intestinal lumen having a monolayer structure on a plate and inner part of an intestinal tract is exposed,
wherein steps (1) and (2) comprise a cell culture medium comprising i) a Wnt agonist, ii) an epidermal growth factor (EGF), iii) a bone morphogenetic protein (BMP) inhibitor and iv) a transforming growth factor- (TGF- β) inhibitor is used, and wherein the Wnt agonist is R-spondin and the BMP inhibitor is noggin.
Allbritton et al. describes a method to generate gastrointestinal epithelial tissue constructs and teaches propagating a gastrointestinal epithelial cell monolayer on said top surface. In some embodiments, the live cells in the monolayer include: (i) undifferentiated cells (e.g., stem or progenitor cells); and (ii) optionally, but in some embodiments preferably, differentiated cells such as enterocytes, Paneth cells,
enteroendocrine cells, tuft cells, microcells, intra-epithelial lymphocytes, and/or goblet cells (See Abstract).
Allbritton et al. teaches the claim 24 (1) by stating that to obtaining the 3D organoid form human intestine and cultured in the 3D organoid model to expand the cells, including stem cells, progenitor cells and differentiated cells (See Example 5; Figure 10 above).
Allbritton et al. teaches 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).
As for the limitation that “…epithelial cells contain differentiated trophoblastic cells…” in claim 24 (2), Allbritton et al. does not explicitly point out the term “trophoblastic cells”. However, Allbritton et al. teaches using a 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).
As for the limitation claim 24 (2) for preparing the 2D organoid from dispersing the 3D organoid to a single cell, Allbritton et al. teaches 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.
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 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). Sato et al. also teaches that a single cell suspension derived from a single-cell-derived-organoid can be replated and grown for 2 weeks (See page 29, lines 20-25) and gastric organoids derived from single cells have been successfully re-plated on a weekly basis for at least 3 months, without losing the properties (See page 74, lines 4-14). Sato et al. also teaches that the paradoxical observation that single cells exposed to a uniform growth-promoting environment can generate asymmetric structures is particularly evident upon scrutiny of the Wnt pathway (See page 45, lines 1-17).
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 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. Therefore, there would have been a reasonable expectation of success given the underlying materials and methods are widely known, successfully demonstrated and commonly used as evidenced by the prior art teachings.
Therefore, the base claim 24 was clearly prima facie obvious to one of ordinary skill in the art at the time the invention.
(New rejection) 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.).
These claims further require that the cell medium or a kit further contains animal-derived bile.
Relevance of Albritton and Sato is set forth above. However, it is silent on the animal-derived bile.
Chang et al. teaches that Bile acids are 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 page 8736, right column, paragraph 1). Chang et al. also teaches that bile acids are as active factors in intestinal content fluid essential for PEC growth (See Abstract), and 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 Allbritton and Sato’s invention 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) Claim 28 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) in view of Sato et al. (WO 2010/090513 A2, published on Aug. 12, 2020) as applied to claims 24 and 27 above, and further in view of Clevers et al. ( US 2014/0243227 Al, published on Aug. 28, 2014).
Regarding the claim 28, it requires the extracellular matrix is basement membrane preparation. Allbritton et al. teaches using the extracellular matrix for support of the organoids, however, it is silent on the basement membrane preparation.
Clevers et al. teaches that alternatively, said ECM is commercially provided. Examples of commercially available extracellular matrices are extracellular matrix proteins (Invitrogen) and basement membrane preparations from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells (e.g. Matrigel™ (BD Biosciences)). (See [0129]).
It would be obvious for one of ordinary skill in the art to select commercial basement membrane preparations as the extracellular matrices for growing the 2D organoid of Albritton.
Responses to Applicant’s Remarks
Applicant’s arguments filed on Feb. 11, 2026 has been received and fully considered.
1). Applicant argued that Allbritton and Sato do not teach or suggest all of the elements of independent claim 1 because neither Albritton nor Sato teaches the use of a medium containing a TGF-β inhibitor (See Remarks, page 6).
The arguments are not persuasive.
In the instant specification, the applicant discloses that the 2D organoid for infection and growth culture of human diarrhea virus according to any one of [2] to [10], in which the TGF-β inhibitor is at least one selected from the group consisting of A83-01, SB-431542, SB-505124, SB-525334, SD-208, LY-36494, and SJN-2511 (See instant specification, e.g. [0020]), where the A83-01 is considered as one of the TGF-β inhibitor. In the teaching of Albritton, Albritton teaches a 2D monolayer culture medium including the A83-01 (500 ng/ml) (See Albritton et al., Example 5, column 20).
2). Applicant argued that Allbritton also teaches the use of ENR-B medium (not containing Wnt-3A) for differentiation into trophoblasts only. Allbritton also teaches medium containing ENR plus a Wnt inhibitor (IWP-2) and a Notch inhibitor (LY-411375) for differentiation into only goblet cells. Hence, the cell culture mediums taught in Allbritton are different from those recited in the pending claims. The addition of Sato does not cure the deficiencies of Allbritton, as Sato also does not teach or suggest the steps of (1) and (2) as recited in independent claim 1 (See Remarks, page 7).
The arguments are not persuasive.
The argument can be summarized in the Figure 5 (See below) for the differentiation of 2D monolayer to specialized cells. Figure 5 teaches that removing the Wnt3A, the stem cell is differentiated to enterocytes cells but not the trophoblasts as argued, and the Goblets cells are formed by adding Wnt-3A, a Wnt agonist. Furthermore, the Example 5 teaches that Human Small Intestinal and Colonic Epithelial Cells can be Cultured as 2D Monolayers and the cell culture medium is the Wnt-3A-conditioned medium (See Allbritton et al., column 20). As for the “Wnt inhibitor” in the argument, Allbritton teaches that the inactivation of both Wnt and Notch signaling enabled differentiation towards goblet cell lineage in the 3D organoid culture model, and no such description in Allbritton for a 2D Monolayers culture. Allbritton did teach testing the inhibitors in the 2D monolayer, but the test is started after 4 days incubation with ENR-WB medium (See Allbritton et al., column 15).
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3). Applicant argued the unexpected effect of producing a 2D organoid being capable of replicating human norovirus (See Remarks, page 7).
The arguments are not persuasive as the follows:
To evaluate if the claimed invention produces unexpected results, one must consider if the results produced by the claimed invention are commensurate in scope with the claims and how the results compare with the closest prior art. See MPEP Section 716.02(d) and (e).
Although the applicant argued the unexpected result for using the claimed 2D organoid for its ability to replicate the human norovirus, and compared the invention with the studies of Takanashi, Papagragko and Herbst-Kralovetz that does not support Norwalk virus (NV) replication in a 3-D INT-407 model (See Remarks, page 8), the claimed unexpected results still did not overcome the teachings of Allbritton and Sato cited in the current office action. Allbritton teaches culturing the 2D Monolayers using the same human cell culture medium starting from a human small intestinal epithelial cell as claimed (See Allbritton, Example 5), which indicates that the cell can inherently to be differentiated to the goblet cells and trophoblastic cells in the 2D organoid.
Applicant argued again that the “Allbritton also teaches the use of ENR-B medium (not containing Wnt-3A) for differentiation into trophoblasts only. Allbritton also teaches medium containing ENR plus a Wnt inhibitor (IWP-2) and a Notch inhibitor (LY-411375) for differentiation into only goblet cells” (See Remarks, page 7). Based on the describes before, there is no evidence to support the arguments in Allbritton’s study. There is no “trophoblasts” differentiation mentioned for removing the Wnt-3, rather than there is an enterocyte differentiation (See Figure 5 above). It is a fact that Allbritton compares the 2D monolayer differentiation under ENR-WB condition (having Wnt) and under the ENR-IG condition (having Wnt inhibitor). Although the result shows that the 2D monolayer cells were successfully differentiated to goblet cells under the ENR-IG condition, the 2D monolayer is cultured under the ENR-WB condition (having Wnt) for 4 days before adding the Wnt inhibitor (See Allbritton, Example 4). Here, the instant claim 1 does not limit to add other reagent to the cell culture medium like Wnt inhibitor. At the same time, Allbritton teaches that the stem cell can be differentiated to goblet cells after the addition of Wnt-3A (See Figure 5 above).
Applicant also argued that the unexpected result based on the instant Example 4 disclosed in the specification. The arguments are not persuasive because the WENRAS medium and ENRA medium in Example 4 are all taught by Allbritton. The meaning of abbreviations are shown below as disclosed in the instant specification:
[0197] W; Wnt3A
[0198] E; EGF
[0199] N; Noggin
[0200] R; Rspondin 1
[0201] A; A-83-01
[0202] S; SB 202190
[0203] L; LY 411575.
D. In addition, as for the phrase “for infection and replication of human norovirus”, it is an “intended use”. Based on MPEP 2111.02, the “intended use” does not add a patentable limitation if the structure of the device is already known in the prior art.
4). The applicant also argued the prior art references cited in the 103 rejections such as Chang and Clevers in claims 13-14, 19, and 27-28.
The arguments are not persuasive.
The references cited in the rejections provide the supports for the teachings of Allbritton and Sato for the specific limitation, for example, Chang’s study indicates that the bile acids derived from porcine is an essential component for enteric virus (including norovirus) replication and growing. Accordingly, these references are applicable to be used as a combination teachings with Allbritton and Sato.
Conclusion
No claims are allowed.
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/RUIXUE WANG/ Examiner, Art Unit 1672
/NICOLE KINSEY WHITE/Primary Examiner, Art Unit 1672