DETAILED ACTION
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Applicant’s response to restriction requirement filed on June 26, 2026 have been received and entered. Claims 1-5, 10, 12-15.19-27, 32 and 34 are pending in the instant application.
Election/Restrictions
Applicant’s election without traverse of claims 1-5, 10, 12-15, 19- 27 (group I) in the reply filed on June 26, 2026 is acknowledged.
Claims 32 and 34 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on June 26, 2026.
Priority
This application is a 371 of PCT/KR2023/001634 filed on 02/06/2023, which claims priority from foreign application no KR 10-2022-0016714 filed on02/09/2022.
Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
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.
Information Disclosure Statement
The information disclosure statements (IDS) submitted on 08/08/2024, 10/07/2025 and 02/23/2026 is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement has been considered by the examiner.
Claims 1-5, 10, 12-15, 19-27 are under consideration.
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.
Claims 1-5, 10, 12-15, 19-26 and 27 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Kwon et al (Sci. Adv. 2021; 7 : eabh1586, 1-17).
Claim recites two operable steps of (a) isolating a pluripotent stem cell-derived three-dimensional intestinal organoid into a single cell or a small cell clump; and (b) two-dimensionally culturing the single cell or the small cell clump in a culture medium comprising a WNT/R-spondin activator, an activator of a prostaglandin signaling pathway, and a receptor tyrosine kinase ligand.
Claim interpretation: The recitation of pluripotent stem cell derived 3D intestinal organoid set forth in claim 1 is produced by a process of claim 15 and therefore phrase is interpreted as product by process (see MPEP 2113). .
With respect to claims 1 and 15, Kwon teaches a method for culturing an intestinal stem cell population, the method comprising
isolating a single cell from a pluripotent stem cell derived 3D-intestinal organoid (see fig. 6A and page 13, col. 1, para. 2), and
culturing the single cell in a culture medium containing B27 supplement, EGF (receptor tyrosine kinase ligand), Noggin (BMP inhibitor), R-spondin1 ( WNT/R-spondin activator), SB202190, prostaglandin E2 (prostaglandin signaling pathway) (se claim page 13, col. 1 and fig. 6a),
Regarding claim 15, Kwon teaches that the pluripotent stem cell-derived three- dimensional intestinal organoid is produced by a method of : (a) culturing human pluripotent stem cells (PSC) in a medium comprising Activin A to produce definitive endoderm (see page 12, col. 2, para. 2);
(b) culturing the definitive endoderm in a medium comprising FGF4, CHIR99021 (GSK3 inhibitors),
(c) culturing the organoid in a culture medium comprising containing DMEM/F12, 2 mM l-glutamine, 2% B27 supplement, EGF, Noggin (BMP inhibitor), R-spondin1, SB202190, prostaglandin E2, nicotinamide to differentiate the definitive endoderm into 3D- HG spheroids; and
(d) culturing the three-dimensional hindgut spheroids in a InS medium comprising: a BMP inhibitor; a Rspondin 1; EGF and any one or more factors selected from the group consisting of IL-2 produce a three-dimensional intestinal organoid (see page 12, col. 2, para. 2 to page 13, col. 1).
With respect to claim 2, Kwon teaches the method, wherein the WNT/R-spondin activator is R-spondin 1 (see page 12, col. 2, and page 13, col. 1).
Regarding claim 3, Kwon teaches that the method, wherein the activator of the prostaglandin signaling pathway is prostaglandin E2 (PGE2) (see page 13, col. 1, para. 2).
With respect to claim 4, Kwon teaches the method, wherein the receptor tyrosine kinase ligand is EGF (see page 13, col. 1, para. 1-2).
Regarding claim 5, Kwon teaches the method, wherein the InS culture medium further comprises B27, N-acetyl-L-cysteine (NAC), nicotinamide, Gastrin, a BMP inhibitor, and a p38 inhibitor (see page 13, col. 1, para. 2).
With respect to claims 10, Kwon teaches the method, wherein the culture medium further comprises Y-27632 (a ROCK inhibitor), and 1 [Symbol font/0x6D]M Jagged-1 (a Notch activator) (see page 13, col. 1, para. 2).
Regarding claims 12-14, Kwon teaches the method, wherein the intestinal stem cell population exhibits an enhanced expression of LGR5, LYZ (see 6C) and Sox9 (fig. 1D). It is relevant to note that Kwon teaches a method step and produces same cell expressing similar markers as one claimed in the instant specification and therefore resulting intestinal stem cell population must necessarily inherently comprise 80% or more of cells comprising an LGR5+ stem cell, and an early enterocyte, as compared to total cells of the population depending on culture duration.
With respect to claim 19-20, the method for producing an intestinal epithelial cell, as disclosed in Kwon, further comprises (a) isolating a pluripotent stem cell-derived three-dimensional intestinal organoid into a single cell or a small cell clump; (b) two-dimensionally culturing the single cell or the small cell clump in a culture medium comprising a WNT/R-spondin activator, an activator of a prostaglandin signaling pathway, and a receptor tyrosine kinase ligand to produce an intestinal stem cell population; and (c) culturing the intestinal stem cell population in a differentiation medium comprising an activator of a prostaglandin signaling pathway, a receptor tyrosine kinase ligand, a p38 inhibitor, an WNT/R-spondin activator, and nicotinamide by trans-well culture system (a air-liquid interface culture system) (see page 13, col. 1, para. 2, wherein the air-liquid interface culture is culturing the intestinal stem cell under a transwell coated with a matrigel (extracellular matrix) (see page 13, col. 1, para. 2).
With respect to claim 21-24, Kwon teaches the method, wherein the WNT/R-spondin activator is a R-spondin 1, the activator of the prostaglandin signaling pathway is any one or more of arachidonic acid (AA), prostaglandin E2 (PGE2), the receptor tyrosine kinase ligand is an epidermal growth factor (EGF), or a basic fibroblast growth factor (bFGF), and p38 inhibitor is SB202190 (see page 13, col. 1, para. 2).
With respect to claims 25-26, Kwon teaches the method, wherein the intestinal epithelial cell exhibits an enhanced expression level for VIL1, LYZ, MUC2 and CDLN3 (see fig. 3 and 6).
Regarding claim 27, Kwon teaches the method, wherein the intestinal epithelial cell comprises a small intestinal cell, a mucus secretory cell, a hormone secretory cell, and a paneth cell (see fig. 1C and 5C).
Accordingly, Kwon anticipates Claims 1-5, 10, 12-15, 19- 27.
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.
Claims 1-5, 10, 12-15, 19-25 and 27 are rejected under 35 U.S.C. 103 as being unpatentable over Koh et al (KR20210119939, dated 100/6/2021), Matsunaga (US 20220033780, filed 10/31/2019) and Clevers et al (WO2012/168930, dated 12/13/2012).
Claim recites two operable steps of (a) isolating a pluripotent stem cell-derived three-dimensional intestinal organoid into a single cell or a small cell clump; and (b) two-dimensionally culturing the single cell or the small cell clump in a culture medium comprising a WNT/R-spondin activator, an activator of a prostaglandin signaling pathway, and a receptor tyrosine kinase ligand.
Claim interpretation: The recitation of pluripotent stem cell derived 3D intestinal organoid set forth in claim 1 is produced by a process of claim 15 and therefore phrase is interpreted as product by process (see MPEP 2113). .
With respect to claims 1-2, 4 and 5, Koh teaches a method comprising (i) isolating a single cell from a 3D- intestinal organoid and (b) culturing the single cells in a culture medium comprising 5% R-spondin conditioned medium (WNT/R-spondin activator), 50 ng/ml EGF {receptor tyrosine kinase) and A83-01 (a TGF-beta inhibitor) for 4 days (see para. 8, 34-38, 40, 43, example 2).
Regarding claim 10, Koh teaches culture medium may further include Rock (Rho-associated, coiled-coil containing protein kinase) inhibitor (see para. 13).
With respect to claim 12, Koh teaches that the intestinal stem cell population exhibits an enhanced expression level for LGR5 (see para. 58, 60).
Koh differs from claimed invention but not disclosing (i) 3D-intestinal organoid is produced from pluripotent stem cell (limitation of claims 1 and 15) and (ii) medium comprises an activator of prostaglandin signaling pathway (limitation of claims 1, 14).
It is noted that method disclosed in Koh explicitly disclose that the origin of the cells producing the organoid is not limited to any specific cells (see para. 8).
Matsunaga cure the deficiency by teaching a method of providing of a 3D-intestinal organoid from a pluripotent stem cell, comprising the following steps (1) to (5): (1) differentiating the pluripotent stem cell into an endoderm-like cell; (2) differentiating the endoderm-like cell obtained in step (1) into an intestinal stem cell-like cell; (3) culturing the intestinal stem cell-like cell obtained in step (2) in the presence of an epidermal growth factor, a fibroblast growth factor, a TGF β receptor inhibitor, a GSK-3 β inhibitor, and a ROCK inhibitor; (4) culturing the cell obtained in step (3) to form a spheroid; and (5) differentiating the spheroid formed in step (4) to form an intestinal organoid, wherein the differentiation comprises culturing in the presence of an epidermal growth factor, a BMP inhibitor, and a Wnt signal activator. It is further disclosed that the plurality of spheroids formed in step (4) are simultaneously cultured in a suspension state, by using a liquid medium adding a material for forming a three-dimensional network structure in an aqueous solution for the culture of step (5) (see claims 1 to 9), wherein the BMP inhibitor is Noggin, and the Wnt signal activator is R-spondin-1 (see ) (limitation of claims 1and 15)
Regarding claim 19, Matsunaga teaches that the method further comprises subjecting the cells constituting the intestinal organoid formed in step (5) to plane culture carried out in the presence of an epidermal growth factor and a TGF β receptor inhibitor, wherein at least part of the plane culture of step (6) is air-liquid interface culture (see .
With respect to claim 20, Matsunaga teaches that the air-liquid interface culture is culturing the intestinal stem cell under a trans-well coated with an extracellular matrix (iMatrix511silk) (see para. 167-168, 209).
The combination of references differs from claimed invention by not disclosing that the (i) differentiation medium also comprises an activator of prostaglandin signaling pathway (claim 1, 3, 19) and the intestinal stem cells express LGR2 and Sox9 (limitation of claim 1 and 12-14).
However, before the effective filing date of instant application in the same technical field, Clevers teaches culturing stem cells and intestinal organoids in a medium containing
R-spondin 1-4 and/or R spondin mimic and one or more inhibitor that directly or indirectly negatively regulates TGF-beta signaling (see claim 1 of ‘930). It is further disclosed that culture medium additionally comprises an activator of the prostaglandin signaling pathway, for example,
PGE2 and/or AA (see claim 14 of ‘210 and page 23) (limitation of claims 1, 3). Clevers further teaches in another embodiment that culture medium comprises cell culture medium comprises advanced-DMEM/F12 preferably supplemented with B27, N2, 200ng/ml N Acetylcysteine, 50ng/ml EGF, 1 μg/ml R-spondin 1-4, 10 nM gastrin, 100 ng/ml FGF10, Nicotinamide, 50ng/ml HGF, 50%a prostaglandin pathway activator, such as PGE2 (50 nM) and/or AA (10 ug/ml) and a TGF-beta inhibitor such as A83-0l (500nM) (see page 42, lines 1-10) and/or P38 inhibitor such as SB202190 (see page 32, lines 10-18 page 4, line 15-16) (limitation of claims 19, 21-24).
With respect to claims 12-13, Clevers teaches intestinal stem cell population exhibits LGR5, CD44 or Sox9 (see page 54, lines 5-7, 14)
With respect to claim 25, Clevers teaches intestinal epithelial cell express VILI (Villin 1), LYZ (Lysozyme), CHGB (Chromogranin B) and MUC2 (Mucin2) (see fig. 7).
Regarding claim 27, Clevers teaches the epithelial cells comprises four differentiated cell types including enterocytes, goblet cells, enteroendocrine cells and Paneth cell (see page 48, lines 12-20).
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art to combine the teachings of prior art to modify the method of Koh by providing pluripotent stem cell-derived three-dimensional intestinal organoid as disclosed in Matsunaga, in the method of culturing an intestinal stem cell population, as instantly claimed, with a reasonable expectation of success, before the effective filing date of the instant invention. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would be motivated to do so because art reported passage culture is considered effective particularly for purifying an intestinal epithelial stem cell-like cell (improvement in purity and uniformity) and more selectively inducing differentiation into the intestinal tract (see para.92) It would have been further obvious to modify the medium of Koh and Matsunaga to further include prostaglandin pathway activator, such as PGE2 as suggested in Clevers (table 2), with a reasonable expectation of success to produce higher number of intestinal stem cells, before the effective filing date of the instant invention. Other limitation of resulting intestinal stem cell population comprising 80% or more of cells comprising a S phase cell, an LGR5+ stem cell, and an early enterocyte, with respect to total cells of the population would be obvious in view of general culture condition and duration disclosed in Matsunaga in view of Clevers that is similar to one disclosed in the instant specification. One of skill in the art would have been expected to have a reasonable expectation of success in culturing an intestinal stem cell population in presence of medium containing PGE2 because prior art successfully reported using PGE2 in the intestinal stem cell expansion medium. It should be noted that the KSR case forecloses the argument that a specific teaching, suggestion, or motivation is required to support a finding of obviousness See the recent Board decision Ex parte Smith, --USPQ2d--, slip op. at 20, (Bd. Pat. App. & Interf. June 25, 2007) (citing KSR, 82 USPQ2d at 1396) (available at http: www. uspto.gov/web/offices/dcom/bpai/prec/fd071925.pdf).
Claims 1 and 26 are rejected under 35 U.S.C. 103 as being unpatentable over Koh et al (KR20210119939, dated 100/6/2021), Matsunaga (US 20220033780, filed 10/31/2019) and Clevers et al (WO2012/168930, dated 12/13/2012) as applied to claim 1 further in view of Traverso et al ( CN110678544, dated 2020-01-10).
The teaching of Koh, Matsunaga and Clevers have been described above and relied in same manner here. The combination of references differs from claimed invention by not disclosing the intestinal epithelial cell exhibits an enhanced expression level for any one or more markers selected from the group consisting of AKR1B15, DHRS11, GALNT4, GALNT5, DHRS3, RDH10, AADAC, NR1I2, SULTEl, DOUX2, FABP1, SLC6A20, SLC43A1, and CLDN3.
Before the effective filing date of instant application, it was routine to characterize intestinal epithelial cells with different markers including FABP1. Traverso teaches intestinal epithelial cells of the intestine could be identified by fatty acid binding protein 1 (FABP1) marker (see claim 16 of ‘544).
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art to combine the respective teachings of Koh, Matsunaga and Clevers to further characterize LGR5 positive intestinal epithelial cell by further looking the expression of FABP1 as suggested in Traverso, with a reasonable expectation of success, before the effective filing date of instant application. A person of skill in the art would have been motivated to characterize implicit characteristics of LGR5 positive intestinal epithelial cell by, as a matter of design choice to further characterize intestinal epithelial cell in order to obtain enriched population of intestinal epithelial cell for transplantation purposes, said design choice amounting to combining prior art elements according to known methods to yield predictable results. One who would have practiced the invention would have had reasonable expectation of success because Traverso had already reported FABP1 is expressed in epithelial cells. It should be noted that the KSR case forecloses the argument that a specific teaching, suggestion, or motivation is required to support a finding of obviousness See the recent Board decision Ex parte Smith, --USPQ2d--, slip op. at 20, (Bd. Pat. App. & Interf. June 25, 2007) (citing KSR, 82 USPQ2d at 1396) (available at http: www. uspto.gov/web/offices/dcom/bpai/prec/fd071925.pdf).
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 1-5, 10, 12-15, 19- 27 are 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.
The term “small cell clump” in claim 1 is a relative term which renders the claim indefinite. The term “small cell clump” 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. It is unclear as to the phrase is small is referred to cell size or cell clump size and how small is a small cell clump. Is it 2 cell , 10, 100 or ore cell clump is small cell clump. Absent any definition, the metes and bounds of the phrase cannot be ascertained. Appropriate correction is required.
Claims 25-27 recite the limitation "intestinal epithelial cell" in line 1. There is insufficient antecedent basis for this limitation in the claim. In the instant case, claim 19 does not produce intestinal epithelial cells and therefore dependent claims 25-27 do not provide antecedent basis for this limitation in the claims. Appropriate correction is required.
Conclusion
No claims allowed.
The prior art made of record and not relied upon is considered pertinent to applicant's disclosure.
Fan et al (Carcinogenesis, 2013, 35, 606-612) teach culturing organoids in presence of exogenous PGE2, PGE3 or dimethyl sulfoxide for 5 days and the number of viable organoids was recorded daily. The results show PGE2 promoted optimal organoid growth and induced significantly higher levels of cell proliferation. It is further disclosed that Lgr5- protein-positive stem cell number is uniquely elevated by >2-fold in PGE2-treated cultures as compared with PGE3 and control (abstract).
Spence (Nature, 2011, 470, 105-110) teaches a pluripotent stem cell derived 3D intestinal organoid produced by a process of culturing pluripotent stem cells in a medium comprising Activin A to differentiate the stem cells to definitive endoderm; (b) culturing the definitive endoderm in a medium comprising: a fibroblast growth factor (FGF) and WNT3A to differentiate the definitive endoderm into three-dimensional hindgut spheroids; and (c) culturing the three-dimensional hindgut spheroids in a medium comprising: noggin (a BMP inhibitor); a R-spondin 1; and EGF (a receptor tyrosine kinase ligand) to produce a three-dimensional intestinal organoid (see page 110, col. 1 under method section).
Sato et al (EP3505620, dated 07/03/2019) teaches a method of producing intestinal stem cells similar to one cited of record.
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/ANOOP K SINGH/Primary Examiner, Art Unit 1632
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