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 .
Applicant's amendments to the claims and arguments filed on 02-04-2026 have been received and entered. Claim 11, 18, 19, 24 have been amended. Claims 1-10, 12-17, 20-23 have been canceled. Claims 25-28 have been added. Claims 11, 18-19, 24-28 are pending in the instant application. The 37 C.F.R. 1.132 declaration by Dr Ito submitted on 02-04-2026 is acknowledged and has been considered in full. The arguments presented therein are discussed below in the Response to Arguments section of this action.
Priority
This application is a CON of PCT/JP2019/029535 filed on 07/26/2019 that claims priority from foreign application JP 2018-141909 filed on 07/27/2018.
The certified English translation of the foreign application JP 2018-141909 submitted on 02-04-2026 is acknowledged.
Claims 11, 18-19, 24-28 are under consideration.
Withdrawn-Claim Rejections - 35 USC § 112
Claims 11, 15, 18-19, 24 were 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. In view of Applicants' amendment of base claim 11, the previous rejections of claims are hereby withdrawn. Applicants' arguments with respect to the withdrawn rejections are thereby rendered moot.
Withdrawn-Claim Rejections - 35 USC§ 103
Claims 11, 15, 19, 24 were rejected under 35 U.S.C. 103 as being unpatentable over Massai et al (Scientific Reports I 7: 3950 I DOI:10.1038/s41598- 017-04158-x, 21 June 2017) in view of Emanuele et al (Pub. No.: US 2016/0002401 Al, Pub. Date: Jan. 7, 2016). In view of Applicants' amendment of base claim 11. Applicants' arguments with respect to the withdrawn rejections are thereby rendered moot. The claims are however subject to new rejections over the prior art of record, as set forth below.
Claim 18 was rejected under 35 U.S.C. 103 as being unpatentable over Massai et al (Scientific Reports I 7: 3950 I DOI:10.1038/s41598- 017-04158-x, 21 June 2017) in view of Emanuele et al (Pub. No.: US 2016/0002401 Al, Pub. Date: Jan. 7, 2016) as applied to claims 11, 15, 19, 24 above, and further in view of Phaechamud et al (Indian J Pharm Sci. 2012 Nov;74(6):498-504. doi: 10.4103/0250-474X.110574.). The rejection is withdrawn for the reasons discussed above.
Claim Objections
Claim 19 and 25 are objected to because of the following informalities:
Claim 19 is clumsy grammatically, making the claim somewhat confusing. The claim reads “wherein the stem cell is suspension cultured by culturing comprises forming cell aggregates”. It is recommended that the phrase be amended to read something like, “wherein the stem cell is suspension cultured such that the culturing comprises forming cell aggregates”.
Claim 25 recites the phrase “selected from poloxamer (Kolliphor P188 BIO) and poloxamer (Kolliphor P407)” which is somewhat confusing because it appears that the terms “Kolliphor P188 BIO” and “Kolliphor P407” in parenthesis are examples species for the genus of poloxamer (see the instant disclosure page 20, lines 1-12). Thus, the claim appears to read on exemplary claim language for the terms “Kolliphor P188 BIO” and “Kolliphor P407”. It would be remedial to delete the parenthesis from each term to read “selected from poloxamer Kolliphor P188 BIO and poloxamer Kolliphor P407”.
Appropriate correction is required.
New-Claim Rejections - 35 USC § 112 - necessitated by amendments
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 11, 18-19, 24-28 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention. This is new matter rejection.
In the instant case, the recitation of “ wherein the proportion of the cultured stem cells that are Oct3/4/SSEA4 positive following the suspension culturing is higher than the proportion of the cultured stem cells that are Oct3/4/SSEA4 positive following culturing in a condition matched control media that does not comprise at least one water-soluble polymer” is considered new matter. Applicants cited [0073] and [0143] of the publication of the instant application for the support of the claimed amendment; however, they do not provide sufficient support for the scope of the claims: Specifically, paragraph [0073] of the instant disclosure only teaches that “The medium of the present invention is preferably provided as a medium for suspension culture of stem cells , more preferably a medium for suspension culture of adult stem cells , embryonic stem cells , and induced pluripotent stem cells, further preferably a medium for suspension culture of embryonic stem cells and induced pluripotent stem cells” ([0073], page 4), and paragraph [0143] only teaches that “The undifferentiated state maintenance rate of cell can be shown by an Oct3/4/SSEA4 positive rate of the cultured Cells” ([0143], page 4).
Upon further review of the instant specification, examiner found that the instant disclosure teaches the study of effect of poloxamer (kolliphor p188 bio) on hiPSC proliferation (Example 3, [0161]-[0164], page 8) that disclose the use of specific Essential 8 medium containing insulin was added 0.1 ug /mL to 1 mg/mL poloxamer (Kolliphor P188 BIO) ([0161], page 8), and 1x106 cells of the 1210B2 line of hiPSC were seeded in a 5 mL bioreactor and cultured with stirring at a stirring rate of 80 rpm as mentioned above . On day 3 , 3.5 mL of the medium was exchanged, the cell aggregates were disrupted on day 4 , and the cell proliferation rate , survival rate and undifferentiated state maintenance rate were determined ([0162], page 8). This working example shows undifferentiation state maintenance rate in Figure 2 (see below). Thus, undifferentiation state maintenance rate that applicants obtained in Figure 2 is tied to a specific cell type of 1210B2 line of hiPSC, a time duration and procedure with Essential 8 medium containing insulin and 0.1 ug /mL to 1 mg/mL Kolliphor P188 BIO poloxamer. However, the claims generically recite the use of any medium and condition with any polyoxyethylene polyoxypropylene block copolymer or polyoxyethylene sorbitan monofatty acid ester with any concentration for any period of time. The guidance provided in the specification is limited to use of a specific medium and specific concentration of the water-soluble polymer. There is no teaching in the instant disclosure that shows using any medium and any concentration of the water-soluble polymer would be suitable for maintenance of undifferentiated state of induced pluripotent stem cell using specific markers of Oct3/4/SSEA4.
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The original claims recite “precipitation of a medium component due to stirring, shaking, circulation or gas bubbling is suppressed” which is entirely different concept than using water-soluble polymer for maintenance of undifferentiated state of induced pluripotent stem cell using specific markers of Oct3/4/SSEA4.
Thus, before the effective filing date of the application was filed, an Artisan of skill in the art would not recognize from the disclosure that Applicant was in possession of using any concentration of water-soluble polymer for maintenance of undifferentiated state of any induced pluripotent stem cell with any medium for any period of time, as claimed. The instant specification does not provide enough guidance for any person skilled in the art to make and use the invention commensurate in scope with the claims.
Claims 18-19, 24-28 directly or indirectly depends from the rejected base claim. This is a new matter rejection
New-Claim Rejections - 35 USC§ 103 - necessitated by amendments
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.
Claims 11, 18-19, 24-25, and 28 are rejected under 35 U.S.C. 103 as being unpatentable over Furue et al (Pub. No.: US 2019/0177686 A1, Foreign Application Priority Data: May 9, 2016) in view of Amit et al (Pub. No.: US 2016/0319241 A1, Pub. Date: Nov. 3, 2016) as evidenced by Kim et al (J Pharm Sci . 2014 Apr;103(4):1043-9. doi: 10.1002/jps.23907. Epub 2014 Feb 15.).
Claim Interpretation
The specification of the claimed invention teaches that poloxamer (polyoxyethylene (160) polyoxypropylene (27) block copolymer) (Kolliphor (registered trade mark) P188 BIO) (BASF) and poloxamer (polyoxyethylene (202) polyoxypropylene (56) block copolymer) (Kolliphor (registered trade mark) P407) (BASF) were used (Examples, page 20- lines 2-6 and Table 1, page 25-line 15- 20). Therefore, poloxamer (Kolliphor P188 BIO) and poloxamer (Kolliphor P407) are interpreted as polyoxyethylene polyoxypropylene block copolymer.
Regarding to claims 11, 24, 25, 28, Furue et al teach method for culturing pluripotent stem cells (title) and culture of pluripotent stem cells capable of maintaining the undifferentiated state and the pluripotency (pluripotent ability ) of the pluripotent stem cells, and a medium used for the culture method of the invention are characterized by containing at least one selected from the group consisting of a Pluronic nonionic surfactant (Abstract). Examples of the pluripotent stem cells include embryonic stem cells (ES cells ), induced pluripotent stem cells (iPS cells ) ([0033], page 3). Examples of the Pluronic nonionic surfactant include Pluronic F-61, Pluronic F-68, Pluronic F-71, and Pluronic F-108 and among these, Pluronic F-68 is preferred ([0037], page 3). It is noted that Pluronic F-68 is Poloxamer 188 (P188) as evidence by Kim et al who teach that Poloxamer 188 (Pluronic F68) is widely used for large-scale production of mammalian cell culture and also where bioreactors are increasingly used to amplify a cell population (Page 1043, right column, 1st para.) (For claim 11, 24-25).
Furue et al teach that using the additive of the invention maintain the undifferentiated state and the pluripotency when an undifferentiation marker such as NANOG OCT3/4, SSEA-3 , SSEA-4 is expressed ([0064], page 5). Furue et al teach when 0.05 mass % Pluronic F-68 was added to the medium, the ratio of the number of cells expressing OCT3/4 to the number of cells stained with DAPI was 74.4 % ([0083], page 6, Example 2), and when culturing pluripotent stein cells in a with a Pluronic nonionic surfactant, the pluripotent stem cells can be proliferated while maintaining the undifferentiated state and the pluripotency of the pluripotent stem cells ([0085], page 6, Example 2) (For claims 11 and 28).
Furthermore, as evidenced by applicant own work, using Poloxamer P188 (Pluronic F-68) in iPSC culture increase Oct3/4/SSEA4 positive iPSC cells as compared with control media that does not comprise water soluble polymer (see Figure 2 of the instant disclosure) (For claim 11).
Furue et al do not teach suspension culturing comprises at least one of shaking, circulation and gas bubbling. Amit et al cure the deficiency.
Amit et al teach well-defined, xeno-free culture media which are capable of maintaining stem cells, and particularly, human embryonic stem cells, in an undifferentiated state are provided (abstract) and a method of culturing pluripotent stem cells in a pluripotent and undifferentiated state, the method comprising culturing the pluripotent stem cells in a suspension culture which comprises a culture medium under conditions devoid of substrate adherence, wherein the pluripotent stem cells are maintained in said suspension culture in a pluripotent and undifferentiated state (see claim 1, page 31). Amit et al teach HA19 medium comprising Pluronic F-68 solution and the F-68 in culture is provided at a concentration of 0.1% ([0244], page 19) and the culturing flasks are gently shaken to suspend cells ([0171], page 13) (For claim 11).
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art before the effective filing date of the rejected claims to combine the teachings of prior art to modify the method of Furue et al by performing culturing the pluripotent stem cells in a suspension culture as taught by Amit et al as instantly claimed, with a reasonable expectation of success. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would have been motivated to do so because Amit et al stated that “The well-defined conditioned media demonstrated in the present study are suitable for culturing hESCs and may be advantageous for undertaking research on the mechanisms of ESC self-maintenance, especially of the possible roles of the LIF/STAT3 pathway and various integrins as fibronectin receptors. Other studies using hESCs, such as the research on differentiation pathways and mechanisms, will benefit from the availability of a well-defined and reproducible culture system” ([0228], page 19), and improvements of the feeder-free, xeno-free culturing systems of hESCs are highly desirable ([0083], page 6). One of ordinary skill in the art would have had a reasonable expectation of success in doing so because Amit et al were successful in culturing and maintaining pluripotent stem cells with working examples and data.
Regarding to claim 18, Furue et al teach high glucose containing DMEM or DMEM /F12 is used as a basal medium, inorganic salts , amino acids, vitamins, saccharides, and minor components are contained , and further, insulin , transferrin , albumin , a lipid group or the like, a growth factor or the like, mercaptoethanol, or the like is added ([0007], page 1). Since Furue et al teach Pluronic F68 (Poloxamer 188) which is the same water-soluble polymer as the claimed invention, it is expected that it has the same function to suppress insulin precipitation.
Regarding to claim 19, Amit et al teach Histological sections of the hESCs clumps formed in the suspension cultures illustrated homogeneous cell population, of round cells with large nucleus (FIGS. 9a-9g) ([0304], page 22), and clumps (i.e., 10-200 cells) are formed ([0134], page 11).
Claims 26-27 are rejected under 35 U.S.C. 103 as being unpatentable over Furue et al (Pub. No.: US 2019/0177686 A1, Foreign Application Priority Data: May 9, 2016) in view of Amit et al (Pub. No.: US 2016/0319241 A1, Pub. Date: Nov. 3, 2016) as evidenced by Kim et al (J Pharm Sci . 2014 Apr;103(4):1043-9. doi: 10.1002/jps.23907. Epub 2014 Feb 15.) as applied to claims11, 18-19, 24-25, and 28 above, and further in view of Elhofy et al (Pub. No.: US 2015/0175956 A1, Pub. Date: Jun. 25, 2015) as evidenced by European Food Safety Authority (Herein EFSA Journal2015;13(7):4152, 74 pp. doi:10.2903/j.efsa.2015.4152.)
The teachings of Furue et al and Amit et al above are incorporated herein in their entirety
Furue et al and Amit et al differ from the claims in that the references fail to disclose polyoxyethylene sorbitan monopalmitate. Elhofy et al cure the deficiency.
Regarding to claims 26-27, Elhofy et al teach media for cell culture (title), and the disclosure provides a media for use with cells in suspension or in adherent culture ([0010], page 1), and the cell is selected from the group consisting of pluripotent stem cells, embryonic stem cells ([0052], page 4). Elhofy et al teach the liquid mix comprises one or more non-ionic surfactants selected from the group consisting of Pluronic-68 (F68 pastille), Pluronic-128, sorbitan, polysorbate and block copolymers. In various embodiments, the non-ionic surfactant is Pluronic-68 (F68 pastille) ([0029], page 3). The polysorbate is polysorbate 20, polysorbate 40, polysorbate 60 or polysorbate 80. Exemplary polysorbates include TWEEN 20 or TWEEN 80 ([0092], page 7). As evidenced by EFSA that teach polyoxyethylene sorbitan monopalmitate is polysorbate 40 (E 434) (see page 3, 1st para, line 4, summary).
Therefore, it would have been prima facie obvious for a person of ordinary skill in the art before the effective filing date of the rejected claims to combine the teachings of prior art to modify the method of Furue et al and Amit et al by using polysorbate 40 (polyoxyethylene sorbitan monopalmitate) in culture medium for suspension culturing of pluripotent stem cells as taught by Elhofy et al as instantly claimed, with a reasonable expectation of success. Said modification amounting to combining prior art elements according to known methods to yield predictable results. One of ordinary skill in the art would have been motivated to do so because Elhofy et al stated that “It is contemplated that media provides advantages to improve cell growth in culture compared to cells cultured not using the serum replacement described herein” (abstract) and “the media compositions or liposome compositions as described herein promote improved culture of cells, e.g., increased cell growth, cell viability or increased expression of recombinant protein, compared to media or liposome compositions comprising a different set of components” ([0059], page 4). One of ordinary skill in the art would have had a reasonable expectation of success in doing so because Elhofy et al were successful in promoting cell growth and viability (see example 2, page 11) with detailed instructions and data.
Response to Arguments
Applicant's arguments filed on 02-04-2025 have been fully considered but they are not persuasive. The claims have been significant amended to change the entire inventive concept from “precipitation of a medium component due to stirring, shaking, circulation or gas bubbling is suppressed” to using “water-soluble polymer” in suspension culture of “embryonic stem cell and induced pluripotent stem cell” to achieve “ wherein the proportion of the cultured stem cells that are Oct3/4/SSEA4 positive following the suspension culturing is higher than the proportion of the cultured stem cells that are Oct3/4/SSEA4 positive following culturing in a condition matched control media that does not comprise at least one water-soluble polymer”. Thus, the new prior arts references necessitated by amendments have been introduced as described above. The Applicant's remarks and the 37 C.F.R. 1.132 declaration by Dr Ito submitted on 02-04-2026 attacking references in preceding office action are thereby rendered moot.
Conclusion
No claim is allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). 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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/KHOA NHAT TRAN/Examiner, Art Unit 1632
/PETER PARAS JR/Supervisory Patent Examiner, Art Unit 1632
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