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 .
Claim Status
1. The amendment filed 11/17/2025 has been entered. Claims 1 – 5, 7 – 10, and 12 – 14 remain pending and are under consideration. Claim 6 has been cancelled.
Priority
2. This application claims priority to U.S. provisional application 63/217,486 filed on July 1, 2021.
Withdrawn Claim Rejections
3. The rejection of claim 6 under 35 U.S.C. 103 is rendered moot in view of Applicant’s cancellation of the claim.
4. The rejection of claims 1 – 5, 7 – 10, and 12 – 14 on the ground of nonstatutory double patenting is withdrawn in view of Applicant’s amendment to the claims and Applicant’s remarks at page 7 – 9.
5. The rejection of claim 6 on the ground of nonstatutory double patenting is rendered moot in view of Applicant’s cancellation of the claim.
Claim Interpretation
4. For the purpose of applying prior art, “wherein the hydrophilic surface allows the pluripotent stem cells to attach and grow” is interpreted as the hydrophilic surface has the capacity for pluripotent stem cells to attach and growth but does not require the attachment and growth of the pluripotent stem cells.
Maintained Claim Rejections
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.
5. Claim(s) 1, 3, 5, 9, 10, 12, and 14 remain rejected under 35 U.S.C. 103 as being unpatentable over Yamada (Yamada R, et. al. J Biosci Bioeng. 2014 Sep;118(3):315-22; previously cited), hereinafter Yamada in view of Ker (Ker, Elmer DF, et al. Biomaterials 32.13 (2011): 3413-3422; previously cited), hereinafter Ker which is cited on the IDS filed 03/23/2023. Although maintained, note that the rejection has been revised in view of the amendment to the claims and cancellation of claim 6.
Regarding claim 1 and 3, Yamada teaches patterned adsorption of gamma globulin alone, vitronectin alone and a combination of gamma globulin and vitronectin (“patterning” of claim 1a; claim 1b; “vitronectin” of claim 3) on a surface and plating pluripotent stem cells on each of the patterned surfaces (claim 1c) (Figure 2A-L; Figure 3 – 5; page 316, left col. last paragraph; page 317, right col. last paragraph; page 319, right col. last paragraph; page 320, left col. paragraph1). Yamada teaches the suitable surface is hydrophilic (“hydrophilic surface” of claim 1) (page 317, left col. last paragraph; Figure 5A; page 321, left col. paragraph 1). Yamada teaches the pluripotent stem cells adhere to the hydrophilic surface and grow (page 319, right col. para. 2; Figure 5). Yamada does not teach “one or a plurality of signaling proteins” of claim 1a.
Regarding claim 9, Yamada teaches patterning gamma globulin in a circle (Figure 3).
Regarding claim 10, Yamada teaches patterning using a PDMS mask (Figure 3; page 316, left col. last paragraph; page 319, right col. paragraph 2).
Regarding claim 14, Yamada teaches human induced pluripotent stem cells (hiPSCs) (page 316, left col. paragraph 4).
Yamada does not teach “one or a plurality of signaling proteins” of claim 1a, concentration of signaling protein of claim 5, or “morphogen” of claim 12. However, Yamada teaches gamma globulin adsorbs to untreated and plasma-treated PDMS surfaces and hiPSCs did not adhere to either surface (page 319, left col. last paragraph). Yamada teaches hiPSCs adhered to the entire surface of PDMS coated with vitronectin alone but did not adhere to the surface coated with gamma globulin alone (page 319, right col. last paragraph; Figure 5). Yamada teaches the advantages of the cell-patterning method include (1) the pattern is long-lasting which is important because hPSC differentiation takes at least a few days; (2) the PDMS sheets can be stored for at least 6 days before cells are plated to enable transportation and quality control, and (3) the patterned surfaces are easy to prepare (page 321, left col. paragraphs 3 – 5). Yamada teaches combination of the patterning method with single-cell dissociation may permit equalization of colony size and thus control of self-renewal and differentiation of hiPSCs (page 321, right col. paragraph 1). Yamada teaches other types of ECM components may be applicable to the method (page 321, right col. paragraph 1). Yamada teaches hiPSCs are a promising source of cells for medical applications and offer the hope of rejection-free transplantation of tissues and patient-specific drug screening (Abstract; page 315, left col. paragraph 1).
Regarding “signaling proteins” of claim 1a and claim 12, Ker teaches solid-phase patterning of BMP2, FGF2, and FGF4 (“signaling proteins” of claim 1a and “morphogen” of claim 12) on fibrin-coated glass coverslips by printing or pipetting followed by cell seeding (page 3, paragraph 4). Ker teaches solid-phase patterning of the signaling proteins spatially directed cell differentiation (page 5, last paragraph; page 6; page 7, paragraph 1 – 2; page 8, last paragraph; page 9, paragraph 3; Figure 3 – 5).
Regarding claim 5, Ker teaches 100 – 200 µg/mL of signaling protein (page 3, paragraph 4). Ker teaches the capability to engineer microenvironmental cues to direct a stem cell population toward multiple fates, simultaneously, in spatially defined regions is important for understanding the maintenance and repair of multi-tissue units (Abstract). Ker teaches extracellular matrix (ECM) can sequester and release growth factors (BMP2, FGF2, FGF4) and can negatively or positively regulate growth factor bioactivity and bioavailability during the maintenance and repair of multi-tissue structures (page 2, paragraph 1). Ker teaches growth factor sequestration by the ECM immobilizes growth factors to specific locations, which in turn imparts the temporal and spatial cues required for directing cell behaviors such as differentiation which is vital for orchestrating complex processes such as development, maintenance and wound healing (page 2, paragraph 1). Ker teaches developing toolsets that can be used to selectively control the physical placement and dosage of multiple exogenous growth factors in a physiologically-relevant manner in order to spatially direct a stem cell population toward multiple cell fates simultaneously is a logical consideration for studying stem cell behaviors and may also have direct applications in regenerative medicine (page 2, paragraph 1).
It would have been obvious prior to the effective filing date of the invention as claimed for the person or ordinary skill in the art to combine the teachings of Yamada regarding a method of patterning a protein and vitronectin by adsorption on a PDMS substrate and plating hiPSCs on the substrate with the teachings of Ker regarding patterning signaling proteins to arrive at the claimed method where BMP2 is patterned on a portion of a PDMS substrate using patterned adsorption; coating the PDMS substrate comprising BMP2 with vitronectin; and plating hiPSCs on the PDMS surface coated with vitronectin. One would have been motivated to combine the teachings of Yamada and Ker in a method to differentiate hiPSCs as Ker teaches developing toolsets that can be used to selectively control the physical placement and dosage of multiple exogenous growth factors in a physiologically-relevant manner in order to spatially direct a stem cell population toward multiple cell fates simultaneously is a logical consideration for studying stem cell behaviors and may also have direct applications in regenerative medicine. One would have a reasonable expectation of success in combining the teachings as Yamada teaches the pattern is long-lasting which is important because hPSC differentiation takes at least a few days and Ker teaches solid-phase patterning of the signaling proteins spatially directed cell differentiation.
6. Claim(s) 2, 4, 7, 8, and 13 remain rejected under 35 U.S.C. 103 as being unpatentable over Yamada (Yamada R, et. al. J Biosci Bioeng. 2014 Sep;118(3):315-22; previously cited), hereinafter Yamada in view of Ker (Ker, Elmer DF, et al. Biomaterials 32.13 (2011): 3413-3422; previously cited), hereinafter Ker which is cited on the IDS filed 03/23/2023 as applied to claims 1, 3, 5, 9, 10, 12, and 14 above, and further in view of Kamp (WO-2011060342-A2; Filed 11/15/2010; Published 05/19/2011; previously cited), hereinafter Kamp.
Yamada in view of Ker make obvious the limitations of claim 1. Ker teaches cells were seeded at 3.1 – 3.6 x 104 cells/cm2 (page 3, paragraph 2) but does not teach about 5 x 105 to about 1 x 106 cells/cm2 of claim 2. Yamada and Ker do not teach a “solubilized basement membrane matrix” of claim 4, “a tissue-culture treated polystyrene plate” of claim 7, “cardiomyocytes” of claim 8, or “BMP4, activin A, noggin, or Wnt3a” of claim 13.
Kamp teaches a matrix overlay-directed cardiogenesis method of iPS cells where 100,000 cells/cm2 (claim 2) are seeded on 6-well tissue culture plates (claim 7) (page 10, 00046 – 00047). Kamp teaches cells were exposed to Activin A, BMP4, (“BMP4” and “activin A” of claim 8) and bFGF with Matrigel (claim 4) overlay (page 10, 00046 – 00047). Kamp teaches the matrix overlay method produced a cell population containing over 90% cardiomyocytes (“cardiomyocytes” of claim 7). Kamp teaches iPS cell-derived cardiomyocytes produced by the matrix overlay method showed organized sarcomere formation and produced large networks of contracting cardiomyocytes with typical cardiac action potentials (page 11, 00050 – 00051). Kamp teaches generating cardiovascular cells from pluripotent stem cells holds great promise for cardiovascular research and therapy and efficient, reproducible methods for differentiating human pluripotent stem cells into cardiovascular cell lineage remain to be elucidated (page 1, 0003 – 0004). Kamp teaches treatment of embryonic stem cells with Activin A and BMP4 promoted cardiogenesis (page 2, 0005). Kamp teaches the method produced cardiomyocytes in high yield (page 3, 00012). Kamp teaches efficient cardiac differentiation of human pluripotent stem cells to cardiomyocytes is effected by simultaneous or sequential application of growth factors such as Activin A, BMP4, and bFGF to pluripotent stem cells grown between layers of matrix (page 4, 00013; page 6, 00030).
It would have been obvious prior to the effective filing date of the invention as claimed for the person of ordinary skill in the art to combine the teachings of Yamada regarding a method of patterning a protein and cell culture substrate by adsorption on a PDMS substrate and plating hiPSCs on the substrate with the teachings of Ker regarding patterning signaling proteins with the teachings of Kamp regarding matrix overlay-directed cardiogenesis method to arrive at the claimed method where BMP4 and Activin A are patterned on a portion of a PDMS substrate using patterned adsorption; coating the PDMS substrate comprising BMP4 and Activin A with Matrigel; and plating hiPSCs on the PDMS surface coated with Matrigel. One would have been motivated to combine the teachings of Yamada, Ker, and Kamp in a method to differentiate hiPSCs to cardiomyocytes as Kamp teaches generating cardiovascular cells from pluripotent stem cells holds great promise for cardiovascular research and therapy and efficient, reproducible methods for differentiating human pluripotent stem cells into cardiovascular cell lineage remain to be elucidated. One would have a reasonable expectation of success in combining the teachings as Ker teaches solid-phase patterning of the signaling proteins spatially directed cell differentiation and Kamp teaches the method produces large networks of contracting cardiomyocytes with typical cardiac action potentials in high yield with sarcomere formation.
Applicant’s Arguments/ Response to Arguments
7. Applicant Argues: On page 5, paragraph 2 and page 6, Applicant asserts that neither Yamada nor Ker teaches patterning of a signaling protein directly on a hydrophilic surface and that a skilled person would be more motivated to plate the signaling protein on a hydrophobic surface such as the surface of Ker or on a vitronectin coated surface like Yamada but not directly on a hydrophilic surface. Applicant asserts that direct binding of a signaling protein to a hydrophilic surface is unpredictable.
Response to Arguments: While Yamada does not teach patterning a signaling protein directly onto a hydrophilic surface, Yamada teaches that gamma-globulin adsorbed onto a plasma-patterned PDMS surface (page 319, right col. para. 2; Figure 3A) and that plasma treatment causes the surface to be hydrophilic as previously cited. Therefore, Yamada teaches that a protein can be patterned on a hydrophilic surface using patterned adsorption (Figure 5). As previously cited, Ker teaches solid-phase patterning of BMP2, FGF2, and FGF4 on fibrin-coated glass coverslips by printing or pipetting followed by cell seeding (page 3, paragraph 4). Ker teaches solid-phase patterning of the signaling proteins spatially directed cell differentiation (page 5, last paragraph; page 6; page 7, paragraph 1 – 2; page 8, last paragraph; page 9, paragraph 3; Figure 3 – 5). Because Yamada teaches that a protein like gamma-globulin which contains a hydrophobic Fc region (page 319, left col. para. 2) can be adsorbed onto a hydrophilic surface, a skilled person would be motivated to plate the signaling proteins of Ker on the hydrophobic surface of Yamada for spatially directed cell differentiation and would have a reasonable expectation of success. Further, because Yamada shows gamma-globulin is adsorbed onto a hydrophilic surface, binding of a signaling protein to a hydrophilic surface it not unpredictable. Additionally, Yamada shows in Figure 5 that both a protein that prefers binding to hydrophobic surfaces (compare left vs. right of panel B in Figure 5) can bind to a hydrophilic surface and that hiPSCs can adhere and grow to the hydrophilic surface with the protein and a cell culture substrate (panel C of Figure 5), binding of a signaling protein to a hydrophilic surface is not unpredictable and one of skill in the art would have a reasonable expectation of success in combining the teachings of Yamada and Ker for spatially directed cell differentiation.
Conclusion
No claims allowed.
THIS ACTION IS MADE FINAL. 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 nonprovisional extension fee (37 CFR 1.17(a)) 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 mailing date of this final action.
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/Z.M.B./Examiner, Art Unit 1632
/MARCIA S NOBLE/Primary Examiner, Art Unit 1632