DETAILED ACTION
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 .
Election/Restrictions
This action is in response to the papers filed on 03/04/2026. Claims 1-22 and 25-29 are currently pending as per claims filed on 11/02/2023. Claims 3, 4, 9, 13-16, 18, 19, 21, and 22 have been amended and claims 23, 24, and 30 have been canceled by Applicants’ amendment filed on 11/02/2023. No claims were added.
Applicant’s election without traverse of Group 1, claims 1-4 and 9-22 in the
reply filed on 03/04/2026 is acknowledged.
Claims 5-8, 25-29 are withdrawn from further consideration by Applicants pursuant to 37 CFR 1.142(b), as being drawn to a nonelected invention, there being no allowable generic or linking claim.
The requirement is still deemed proper and is therefore made FINAL.
Therefore, claims 1-4 and 9-22 are subject to examination to which the
following grounds of rejection are applicable.
Priority
The instant application claims domestic benefit to US provisional patent
application number 63/390,458 filed on 07/19/2022. Thus, the earliest possible priority
for the instant application is 07/19/2022.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on 02/02/2024 was filed after the mailing date of the current office action. The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
It is noted that references U.S. Appl. No. 18/354,212, 18/354,210, and 18/354,201 listed in the IDS correspond to U.S. PG Publication No. 20240132851 ,20240052319, and 20240052318, respectively.
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 2, 3, 10-15, 17, 18, and 20 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.
Claim 2 is vague and indefinite in the recitation of “culturing the cells of (ii) in a base culture medium before (iii)”. It is unclear if the cells of (ii) are in the same base culture medium of step (ii) of claim 1, or if there is an alternative base culture medium for the cells of (ii) before being cultured in step (iii). As such the metes and bounds of the claim are indefinite.
Claim 3 is vague and indefinite in the recitation of “culturing the cells of (iii) in a base culture medium”. It is unclear if the cells of (iii) are in the same base culture medium of step (iii) of claim 1, or if there is an alternative base culture medium for the cells of (iii). As such the metes and bounds of the claim are indefinite.
Claims 10, 12-15, 17, 18, and 20, recites the limitation "the culture medium". There is insufficient antecedent basis for this limitation in the claim.
Claims 1-4 recites the limitation "the cells" (line 7, 9, 11, 14, and 16). There is insufficient antecedent basis for this limitation in the claim.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claim(s) 1-4 and 9-22 are rejected under 35 U.S.C. 103 as being unpatentable over Orlova et al (Nature Protocols, 2014, pages 1514-1531; as cited in IDS), and further in view of Watanabe et al (Nature Biotechnology, 2007, pages 681-686).
Regarding claim 1, Orlova teaches a method of differentiating human pluripotent stem cells (hPSCs) into pericytes (i.e. a species of stromal cells) comprising:
Culturing hPSCs to induce mesoderm specification by the addition of bone morphogenetic protein 4 (BMP4), activin A, small-molecule inhibitor of glycogen synthase kinase-3β (CHIR), and vascular endothelial growth factor (VEGF) (page 1516, left col, para 1) to a base culture medium (B(P)EL medium (page 1521, Mesoderm induction medium; page 1523, step 13 – Monolayer differentiation of hPSCs));
Removal of mesoderm-inductive factors on day 3 of differentiation and replace with vascular specification medium (base culture medium (B(P)EL medium supplemented with VEGF and the transforming growth factor-β (TGF-β) pathway small-molecule inhibitor SB431542) (page 1521, vascular specification medium; page 1516, left col, para 1);
Cell sorting to isolate endothelial cells (ECs) fractions from pericytes fraction by Dynabead isolation; and
Passage of pericyte cell fraction into DMEM–10% (vol/vol) FBS (base culture medium) with TGF-β3 and PDGF-BB and incubate for 3 days to expand pericytes (page 1524, Expansion of pericytes: step (v))
rendering obvious a method for differentiating pluripotent stem cells into stromal cells, comprising:
culturing pluripotent stem cells in a base culture medium comprising bone morphogenetic protein 4 (BMP4), vascular endothelial growth factor (VEGF), a glycogen synthase kinase 3 (GSK3) inhibitor, activin A.
culturing the cells of (i) in a base culture medium comprising VEGF and a transforming growth factor β (TGFβ) inhibitor;
culturing the cells of (ii) in a base culture medium comprising TGFβ3 and a platelet- derived growth factor (PDGF) to form stromal cells.
Orlova does not teach Rho-associated, coiled-coil containing protein kinase (ROCK) inhibitor in step (i).
However, Watanabe teaches the use of the ROCK inhibitor Y-27632 enables “human embryonic stem cells to grow and differentiate under unfavorable culture conditions” (page 684, right col, para 3; page 685, left col, para 1).
It would have been prima facie obvious to one of ordinary skill, in the art at the time of the effective filing date, to modify the teachings of a method of differentiating pluripotent stem cells into pericytes (i.e. a species of stromal cells) from Orlova in the culture step (I) to further add the ROCK inhibitor from Watanabe to increase the survival of the pluripotent cells undergoing differentiation as the process of differentiation is stressful to cells and the addition of ROCK inhibitor would mitigate cell loss during culture. One would be motivated to do so to maximize cell survival and, therefore, maximize the number of differentiating cells in the culture. As use of ROCK inhibitor to increase cell viability during cell culture is known in the art, one would have a reasonable expectation of success.
Regarding claim 2, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches the cells of step (ii) are cultured in vascular specification medium which has b(P)EL base medium (page 1521 - table for composition), rendering obvious culturing the cells of (ii) in a base culture medium before (iii).
Regarding claim 3, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches the cells of step (iv) are then cultured with DMEM–10% (vol/vol) FBS (base culture medium) at step (v) (page 1524, Expanding pericytes step (v) and (vi)), rendering obvious step (iv) culturing the cells of (iii) in a base culture medium.
Regarding claim 4, the teachings of Orlova and Watanabe render obvious claim 1. In particular, Orlova discloses culture of hPSCs in a vascular specification medium (base culture medium (B(P)EL medium supplemented with VEGF and the transforming growth factor-β (TGF-β) pathway small-molecule inhibitor SB431542) (page 1521, vascular specification medium; page 1516, left col, para 1) ; Though Orlova, does not teach that cells cultured in VEGF and TGF-β are not passaged before and additional culture step (iii) comprising at least PDGF, it would have been prima facie obvious for one of ordinary skill in the art not to passage the cells based on influential considerations in the design of the culture assay such as cell confluency, growth factors and inhibitors present in the media, cell differentiation and other considerations.
Regarding claim 9, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Watanabe teaches using the ROCK inhibitor Y-27632 (page 684, right col, para 3), rendering obvious wherein the ROCK inhibitor is Y-27632.
Regarding claim 10, the teachings of Orlova and Watanabe render obvious claim 1 and 9. Moreover, Watanabe teaches that 10 μm is a commonly used concentration for Y-27632 in cell culture (page 681, right col, para 2), rendering obvious wherein Y-27632 is present in the culture medium at a concentration of about 10 μM..
Regarding claim 11, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches using the small-molecule inhibitor of glycogen synthase kinase-3β (CHIR) (page 1516, left col, para 1), rendering obvious wherein the GSK3 inhibitor is CHIR99021.
Regarding claim 12, the teachings of Orlova and Watanabe render obvious claim 1 and 11. Moreover, Orlova teaches that the mesoderm induction media contains 1.5 μm CHIR (page 1521, mesoderm induction media), rendering obvious wherein CHIR99021 is present in the culture medium at a concentration of about 1.5μM.
Regarding claim 13, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches that the mesoderm induction media contains 30 ng/ml BMP4 (page 1521, mesoderm induction media), rendering obvious wherein BMP4 is present in the culture medium at a concentration of about 30 ng/mL.
Regarding claim 14, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches that the mesoderm induction media contains 50 ng/ml VEGF (page 1521, mesoderm induction media), rendering obvious wherein VEGF is present in the culture medium at a concentration of about 50 ng/mL.
Regarding claim 15, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches that the mesoderm induction media contains 25 ng/ml Activin A (page 1521, mesoderm induction media), rendering obvious wherein activin A is present in the culture medium at a concentration of about 25 ng/mL.
Regarding claim 16, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches the transforming growth factor-β (TGF-β) pathway small-molecule inhibitor is SB431542 (page 1516, left col, para 1; page 1521, vascular specification media), rendering obvious wherein the TGF3 inhibitor is SB431542.
Regarding claim 17, the teachings of Orlova and Watanabe render obvious claim 1 and 16. Moreover, Orlova teaches the transforming growth factor-β (TGF-β) pathway small-molecule inhibitor SB431542 has a concentration of 10 μM (page 1521, vascular specification media), rendering obvious wherein SB431542 is present in the culture medium at a concentration of about 10 μM.
Regarding claim 18, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches the cells are passaged and incubated with media containing 2 ng/mL of TGF-β3 (page 1524, Expanding pericytes step (v)), rendering obvious wherein TGFβ3 is present in the culture medium at a concentration of about 2 ng/mL.
Regarding claim 19, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches the cells are passaged and incubated with media containing PDGF-BB (page 1524, Expanding pericytes step (v)), rendering obvious wherein the PDGF is PDGF-BB.
Regarding claim 20, the teachings of Orlova and Watanabe render obvious claim 1 and 19. Moreover, Orlova teaches the cells are passaged and incubated with media containing 4 ng/mL of PDGF-BB (page 1524, Expanding pericytes step (v)), rendering obvious wherein PDGF-BB is present in the culture medium at a concentration of about 4 ng/mL.
Regarding claim 21, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches a protocol for the generation of pericytes from human pluripotent stem cells (abstract, page 1), rendering obvious wherein the stromal cells are pericytes.
Regarding claim 22, the teachings of Orlova and Watanabe render obvious claim 1. Moreover, Orlova teaches not fully confluent cells at Day 0 (Figure 2D shows Day 0 cells that are not fully confluent), rendering obvious wherein the pluripotent stem cells are not fully confluent at the start of the culturing of (i).
Conclusion
Claims 1-4 and 9-22 are rejected.
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/JULIANA IRENE CANDELARIA/ Examiner, Art Unit 1634
/MARIA G LEAVITT/ Supervisory Patent Examiner, Art Unit 1634
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