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 .
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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 24 October 2025 has been entered.
Claims 1, 3-11, 19, 20, and 23-26 have been amended. Claims 11-20 remain withdrawn. Claims 1, 3-10, and 23-26 are currently pending and under examination.
This application is a U.S. National Phase Application under 35 U.S.C. §371 of International Patent Application No. PCT/SG2020/050339, filed June 18, 2020, and claims priority to Singapore Application No. 10201905610Y, filed June 18, 2019.
Withdrawal of Objections/Rejections:
The rejection of claims 1, 3-10, and 23-26 under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite, is withdrawn.
The rejection of claim 3 under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, as being of improper dependent form, is withdrawn.
Maintenance/Modification of Objections/Rejections:
Claim Objections
Claim 23 is objected to because of the following informalities: “of” should be placed in front of “0.5µM” on line 2 (i.e. “at a concentration of 0.5 µM”). Appropriate correction is required.
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, 3-10, and 23-26 are rejected under 35 U.S.C. 103 as being unpatentable over Hanna et al. (US 2017/0275593; Published 2017 – Previously Presented), in view of Kuppers-Munther (IDS; US 2019/0024044, Published Jan. 24, 2019).
With regard to claim 1, Hanna et al. teach a cell culture medium to generate and expand pluripotent stem cells, which is maintaining or increasing pluripotency in a cell cultured in the medium, the medium comprising components including a GSK3 inhibitor, including CHIR99021 or Kenpaullone, in an amount including between 0.5-10 µM; and an SRC family kinase inhibitor, including Dasatinib, in an amount including between about 0.5-10 µM (Abs.; Para. 1, 16, 95-119, 330, 361-362). It would have been obvious to an ordinary artisan to utilize an amount of GSK3 inhibitor and Dasatinib within the range expressly taught by Hanna et al., which fully encompasses a concentration of 0.5 GSK3 inhibitor and 0.5 Dasatinib. Hanna et al. additionally teach that culturing is performed under culture conditions devoid of feeder cells (Para. 82); a such, the cell culture medium is a feeder-free cell culture medium.
Hanna et al. do not specifically teach that the GSK3 inhibitor is AZD5438.
Kuppers-Munther teaches that GSK3 inhibitors usable in cell culture media include CHIR99021, Kenpaullone, and AZD5438 (Para. 166).
It would have been obvious to one of ordinary skill in the art to combine the teachings of Hanna et al. and Kuppers-Munther, because both teach GSK3 inhibitors that are usable in cell culture media, including CHIR99021 and Kenpaullone. The use of AZD5438 as a GSK3 inhibitor in cell culture media is known in the art as taught by Kuppers-Munther. The use of AZD5438 as taught by Kuppers-Munther in place of CHIR99021 or Kenpaullone in the cell culture medium of Hanna et al. amounts to the simple substitution of one known GSK3 inhibitor for another, and would have been expected to predictably and successfully provide an alternative GSK3 inhibitor for use in the cell culture medium.
It is again noted that the teachings of Hanna et al. of a cell culture medium to generate and expand pluripotent stem cells, is deemed to be maintaining or increasing pluripotency. Further, taken together, Hanna et al. and Kuppers-Munther render obvious the feeder-free cell culture medium as claimed, including all components as claimed. A chemical composition and its properties are inseparable (see MPEP 2112.01, II). As the medium cannot be separated from its properties, the medium as rendered obvious would necessarily be capable of maintaining or increasing pluripotency in a cell cultured in the feeder-free cell culture medium when used.
With regard to claim 3, Hanna et al. teach that the GSK3 inhibitor is present in an amount including between 0.1-50 µM; and the SRC family kinase inhibitor, including Dasatinib, is present in an amount including between 0.1-70 µM (Para. 330-331, 361-362). It would have been obvious to an ordinary artisan to utilize an amount of the GSK3 inhibitor and Dasatinib within the range expressly taught by Hanna et al., which fully encompasses concentrations of 0.1 µM GSK3 inhibitor, which includes AZD5438 as rendered obvious by Kuppers-Munther, and 0.1µM Dasatinib.
With regard to claim 4, Hanna et al. teach that the feeder-free cell culture medium may further comprise SB590885, including at a concentration of 0.25-0.5 µM (Para. 398; Fig. 18), which is fully encompassed within 0.1 to 2.5 µM.
The feeder-free cell culture medium may further comprise PD0325901, including at a concentration of about 1 µM (Para. 307, 456; Fig. 17, 19), which is fully encompassed within 0.2 to 10 µM.
The feeder-free cell culture medium may further comprise Y-27632, including at a concentration of about 5 µM (Para. 337-338), which is fully encompassed within 5 to 20 µM.
As Hanna et al. expressly teach that SB590885, PD0325901, and/or Y-27632 may be present in the cell culture medium, it would have been obvious to an ordinary artisan to utilize one or more of these taught components in the cell culture medium.
With regard to claim 5, Hanna et al. teach that the feeder-free cell culture medium further comprises: between about 1-1000 ng/ml (.001-1 µg/ml) LIF, including recombinant human LIF (Para. 297, 520-521), wherein it would have been routine for an ordinary artisan to optimize the amount of LIF present in the culture medium, dependent on the cell type to be cultured and the culture conditions, including providing 5-20 µg/ml LIF; between about 0.2-10 µM PD0325901 (Para. 307), which is fully encompassed within 0.2 to 10 µM; between about 0.1-2 µM, or about 0.5 µM, SB590885 (Para. 398), which are fully encompassed within 0.1 to 2.5 µM; between about 1-3 µM, or about 1.5 µM, Src family kinase inhibitor, including WH4-023 (Para. 359-362), which are fully encompassed within 0.1 to 2.5 µM; about 5 µM Y-27632 (Para. 337-338), which is fully encompassed within 5 to 20 µM; and about 20 ng/ml Activin A (Para. 17, 424), which is fully encompassed within 5 to 20 mg/ml.
With regard to claim 6, Hanna et al. teach that the feeder-free cell culture medium comprises: DMEM/F12, Neurobasal medium, N2, B27, 2 mM L-glutamine, 5 mL non-essential amino acids, 50 µl of 50 mM β-mercaptoethanol, 5 mL penicillin-streptomycin, 100x Fraction V 7.5% solution BSA, 20 ng/ml LIF, 1 mM PD0325901, and 10 mM Y-27632 (Ex. 1; Para. 914-919, Para. 398).
It is further taught that the feeder-free cell culture medium can comprise between about 1-1000 ng/ml (.001-1 µg/ml) LIF, including recombinant human LIF (Para. 297, 520-521), wherein it would have been routine for an ordinary artisan to optimize the amount of LIF present in the culture medium, dependent on the cell type to be cultured and the culture conditions, including providing 10 µg/ml LIF. Additionally, the feeder-free cell culture medium can comprise between about 1-3 µM, or about 1.5 µM, Src family kinase inhibitor, including WH4-023 (Para. 359-362), which fully encompass 1 µM. Further, the feeder-free cell culture medium can comprise about 20 ng/ml Activin A (Para. 17, 424), wherein it would have been routine for an ordinary artisan to optimize the amount of Activin A present in the culture medium, dependent on the cell type to be cultured and the culture conditions, including providing 10 ng/ml Activin A.
Additionally, it would have been routine for an ordinary artisan to optimize the concentration of taught components within the feeder-free cell culture medium including the non-essential amino acids, β-mercaptoethanol, penicillin-streptomycin, BSA, LIF, and Activin A, depending on the cell type to be cultured and the culture conditions. Further, it noted that "the discovery of an optimum value of a variable in a known process is usually obvious." Pfizer v. Apotex, 480 F.3d at 1368. The rationale for determining the optimal parameters for prior art result effective variables "flows from the 'normal desire of scientists or artisans to improve upon what is already generally known.'" Id. (quoting In re Peterson, 315 F.3d 1325, 1330 (Fed. Cir. 2003)). Accordingly, it would have been obvious to optimize the amounts of the taught components in the feeder-free cell culture medium, including to 1% non-essential amino acids, 0.1 mM β-mercaptoethanol, 1% penicillin-streptomycin, 50 µg/ml BSA,10 µg/ml LIF, and 10 ng/ml Activin A, to result in a cell culture medium with an effective amount of each taught component to provide the optimal growth environment for the desired cell type(s) being cultured.
With regard to claims 7-10 and 26, taken together, Hanna et al. and Kuppers-Munther render obvious the feeder-free culture medium as claimed, including the components as claimed. As the feeder-free culture medium cannot be separated from its properties, the feeder-free culture medium as rendered obvious by Hanna et al. and Kuppers-Munther necessarily is capable of: maintaining or increasing pluripotency in a cell cultured in the cell culture medium in the absence of co-culture such as feeder cells, wherein the pluripotency comprises expression of a naive pluripotent stem cell marker selected from the group consisting of: CD130, CD75, NMT3L, DPPA5, KLF5, TFCP2L1, KLF4, DPPA3, NANOG, KLF17, POU5F1, and PRDM14; maintaining or increasing pluripotency in a cell cultured for 5 or more passages; and decreasing the expression of a primed pluripotent stem cell marker, including ZIC2 or B3GAT1, in a cell cultured in the cell culture medium.
With regard to claims 23 and 24, Hanna et al. teach that the feeder-free cell culture medium may further comprise: SB590885, including at a concentration of 0.5 µM (Para. 398; Fig. 18); PD0325901, including at a concentration of 1 µM (Para. 307, 456, 917); Y-27632, including at a concentration of 10 µM (Para. 337-338, 917); and Activin A, including at a concentration from about 1-40 ng/ml (Para. 424), wherein it would have been obvious to an ordinary artisan to utilize an amount within the expressly taught range, including 10 ng/ml.
With regard to claims 25, Hanna et al. teach a 1:1 ratio of F12 DMEM and Neurobasal medium, and N2 supplement, B27 supplement, L-Glutamine, and non-essential amino acids (Para. 289-291, 599).
Hanna et al. further teach that the feeder-free cell culture medium comprises: 50 µl of 50 mM β-mercaptoethanol and 100x Fraction V 7.5% solution BSA (Ex. 1; Para. 914-919, Para. 398). It would have been routine for an ordinary artisan to optimize the concentration of taught components within the cell culture medium, including the β-mercaptoethanol and BSA, depending on the cell type to be cultured and the culture conditions. Further, it noted that "the discovery of an optimum value of a variable in a known process is usually obvious." Pfizer v. Apotex, 480 F.3d at 1368. The rationale for determining the optimal parameters for prior art result effective variables "flows from the 'normal desire of scientists or artisans to improve upon what is already generally known.'" Id. (quoting In re Peterson, 315 F.3d 1325, 1330 (Fed. Cir. 2003)). Accordingly, it would have been obvious to optimize the amounts of the taught components in the cell culture medium, including 0.1 mM β-mercaptoethanol and 62.5 ng/ml BSA, to result in a cell culture medium with an effective amount of each taught component to provide the optimal growth environment for the desired cell type(s) being cultured.
Response to Arguments
Applicant urges that Hanna in view of Kuppers-Munther do not teach that the culture medium is capable of maintaining or increasing pluripotency. Additionally, the cell culture medium of Hanna requires a STAT3 activator, an ERK1/2 inhibitor, and an Axin stabilizer, while the current invention does not. Additionally, there is no motivation to modify Hanna, including using Kuppers-Munther, to eliminate the STAT3 activator, ERK1/2 inhibitor, and Axin stabilizer in Hanna’s medium and substitute AZD5438 and Dasatinib. Further, one would not have been motivated to combine the teachings of Hanna with Kuppers-Munther to obtain the claimed medium, as Hanna teaches culturing pluripotent stem cells, while Kuppers-Munther’s method teaches a differentiation medium to obtain mature hepatocytes.
Applicant’s arguments have been fully considered, but have not been found persuasive.
With regard to Applicant’s argument that Hanna in view of Kuppers-Munther do not teach that the culture medium is capable of maintaining or increasing pluripotency; as noted previously, the teachings of Hanna et al., of a cell culture medium to generate and expand pluripotent stem cells, is deemed to be maintaining or increasing pluripotency. Further, taken together, Hanna et al. and Kuppers-Munther render obvious the feeder-free cell culture medium as claimed, including all components as claimed. A chemical composition and its properties are inseparable (see MPEP 2112.01, II). As the medium cannot be separated from its properties, the medium as rendered obvious would necessarily be capable of maintaining or increasing pluripotency in a cell cultured in the feeder-free cell culture medium when used.
With regard to Applicant’s argument that the cell culture medium of Hanna requires a STAT3 activator, and ERK1/2 inhibitor, and an Axin stabilizer, while the current invention does not, and there is no motivation to modify Hanna, including using Kuppers-Munther, to eliminate the STAT3 activator, ERK1/2 inhibitor, and Axin stabilizer in Hanna’s medium and substitute AZD5438 and Dasatinib; it is noted that Applicant utilizes the transitional phrase “comprising.” The transitional phrase "comprising" is inclusive or open-ended and does not exclude additional, unrecited elements or method steps (see MPEP 2111.03, I.). As such, the presence of additional components including a STAT3 activator, ERK1/2 inhibitor, and Axin stabilizer in Hanna’s medium is permissible. Should Applicant wish to exclude any additional, unrecited component, the transitional phrase “consisting of” should be utilized.
With regard to Applicant’s argument that one would not have been motivated to combine the teachings of Hanna with Kuppers-Munther to obtain the claimed medium, as Hanna teaches culturing pluripotent stem cells, while Kuppers-Munther’s method teaches a differentiation medium to obtain mature hepatocytes; as noted previously, it would have been obvious to one of ordinary skill in the art to combine the teachings of Hanna et al. and Kuppers-Munther, because both teach GSK3 inhibitors that are usable in cell culture media, including CHIR99021 and Kenpaullone. The use of AZD5438 as a GSK3 inhibitor in cell culture media is known in the art as taught by Kuppers-Munther. The use of AZD5438 as taught by Kuppers-Munther in place of CHIR99021 or Kenpaullone in the cell culture medium of Hanna et al. amounts to the simple substitution of one known GSK3 inhibitor for another, and would have been expected to predictably and successfully provide an alternative GSK3 inhibitor for use in the cell culture medium.
Conclusion
No claims are allowable.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to JENNIFER M.H. TICHY whose telephone number is (571)272-3274. The examiner can normally be reached Monday-Thursday, 9:00am-7:00pm ET.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Sharmila G. Landau can be reached at (571)272-0614. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/JENNIFER M.H. TICHY/Primary Examiner, Art Unit 1653