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 .
Claim status
On 16 January 2026, Applicant has amended claims 1-4, 6-9, 11-20, 22 and 24. Claims 5, 10, 21, 23 and 25-28 have been cancelled and added new Claims 29-31. Claims 1-4, 6-9, 11-20, 22, 24 and 29-31 are pending.
Therefore, claims 1-4, 6-9, 11-20, 22, 24 and 29-31 are under current examination.
Priority
This application was filed 01/20/2023 and is a 371 application of PCT/NL2021/050474 filed on 07/23/2021, claims, filed 07/23/2020, which claims benefit to the foreign application 2026121 filed on 07/23/2020. Thus, the earliest possible priority for the instant application is 07/23/2020.
Withdrawn Claim Objections
The objection to Claim 22 has been withdrawn due to applicant’s amendment.
Withdrawn Abstract Objections
Applicant has amended the Specification to include the abstract on its own page in compliance with MPEP 608.01(b). Therefore, the objection to abstract has been withdrawn.
Withdrawn Rejections
In the remark filed on 16 January 2026, applicant discloses that “the specification provides an express, detailed, and enabling definition of "preselected cell type." In particular, the Specification states: "As used herein, 'preselected cell type' refers to a cell of a certain type that was preselected as the cell type to be obtained with the method as disclosed herein ... " (Specification, pg. 7 line 31 through pg. 8 line 23)”. Therefore, the Applicant’s arguments have been fully considered and persuasive. Accordingly, the prior rejection of claims 1-4, 6-9, 11-15, 17-20, 22 and 24 under 35 U.S.C. 112(b) is withdrawn.
In the reply filed 16 January 2026, applicant amended the claim 1 and exclusively impose a closed culture system for the in vitro manufacture of a preselected-cell type differentiated from a pluripotent stem cell culture method. In the prior art prior art Kempf’s methods, including bioreactor-based methods, require steps like centrifugation, filtration, and/or manual wash steps. Accordingly, Kempf fails to disclose a closed culture system for the in vitro manufacture of a preselected-cell type as required for amended claims. Therefore, the prior rejection of claims 1-4, 6-9, 11-18, 20, and 24 under 35 U.S.C. 102(a)(1) as being anticipated by Kempf et al., (Nature Protocols, 10(9), 2015, pp. 1345-1361; cited in IDS filed 01/20/2023; hereinafter “Kempf”) is withdrawn.
Consequently, the prior rejection of claims 1-4, 6-9, 11-20, 22 and 24 under 35 U.S.C. 103 as being unpatentable over Kempf et al., in view of Nishimura et al., (Experimental hematology, 80, pp.16-20; 2019; cited in PTO892; hereinafter “Nishimura”), further in view of Arauchi et al. (Frontiers in Endocrinology, 8, p.103, 2017; cited in PTO892; hereinafter “Arauchi”) is withdrawn.
New 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.
Claim 14 is 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.
Dependent claim 14 is recited a limitation “the compositions of the different culture media”. However, the independent claim 1 does not introduce this limitation, therefore there is insufficient antecedent basis for this limitation. Appropriate correction is required.
New 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 of this title, 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 set forth in Graham v. John Deere Co., 383 U.S. 1, 148 USPQ 459 (1966), that are applied 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.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-4, 6-9, 11-13, 15-20, 22, 24 and 29-31 are rejected in modified form under 35 U.S.C. 103 as being unpatentable over Timmins et al., (US20190031990A1; pub. Date: Jan. 31 2019; cited in PTO892; hereinafter “Timmins”), in view of Burridge et al., (PloS one, 6(4), p.e18293, 2011; cited in PTO892; hereinafter “Burridge”), further in view of Arauchi et al. (Frontiers in Endocrinology, 8, p.103, 2017; cited in PTO892; hereinafter “Arauchi”). The new rejection is necessitated by applicant’s claim amendments.
With respect to claims 1,16 and 24, Timmins teaches a method for the in vitro (e.g., in a bioreactor, MiniBio 500 STR system [0162]) manufacture of a preselected cell type (i.e., Cardiomyocytes (CMs) [0058], [0170]) differentiated from a pluripotent stem cell ([0058], [0170]), in a closed culture system ([0012], [0037], [0042], [0050] of Timmins), wherein the method comprises the steps of:
providing pluripotent stem cells and a culture medium ([0007], [0033], [0053] of Timmins);
introducing the pluripotent stem cells and the culture medium into a culture vessel ([0058], [0170], [0046]), wherein the culture vessel is part of a closed culture system ([0012], [0037], [0042], [0050] of Timmins), wherein the culture medium is
a culture medium for proliferation of the pluripotent stem cells (mTeSR1 with Y27632 is a culture medium for proliferation ([0024], [0141] of Timmins); or
a culture medium for inducing differentiation of the pluripotent stem cells (Example 4, [0168]-[0170] of Timmins);
mixing the culture medium in the culture vessel thereby allowing the cells to grow in the form of cell aggregates (e.g., cardiomyocytes [0128] of Timmins) and preventing settling of the cell aggregates ([0039], [0141] of Timmins);
discontinuing the mixing of the culture medium in the culture vessel thereby allowing the cell aggregates to settle ([0128] of Timmins);
collecting part of the culture medium in the culture vessel ([0128] of Timmins);
optionally, in case in step b) a culture medium for proliferation of the pluripotent stem cells was used, introducing a further culture medium for proliferation of the pluripotent stem cells ([0141]) in the culture vessel and repeating step c) - e);
introducing a subsequent culture medium into the culture vessel, wherein the culture medium is a culture medium for inducing differentiation of the cells towards the preselected cell type ([0171] of Timmins);
mixing the culture medium in the culture vessel thereby allowing the cells to grow in the form of cell aggregates ([0077], [0095] of Timmins). Although Timmins doesn’t mention that mixing the culture medium can preventing settling of the cell aggregates, however, Timmins teaches the a) geometry of the reactor vessel; (b) mixing mechanism (e.g., impeller type); (c) duration of the dissociation process could impact on the choice of reactor. Therefore, the geometry and mixing mechanism may be based on requirements for cultivating cells and POSITA would recognize that mixing the culture medium can preventing settling of the cell aggregates.
Timmins teaches that the bioreactors were equipped with marine impellers. discontinuing the mixing of the culture medium in the culture vessel thereby allowing the cell aggregates to settle ([0141], see MPEP 2144.04 (II) B);
collecting the culture medium in the culture vessel, collecting the cell aggregates in the culture vessel, or collecting both ([0127]-[0128] of Timmins).
Nevertheless, it would have been obvious to one having ordinary skill in the art at the time the invention was filed to the in vitro manufacturing method of a preselected cell type (i.e., Cardiomyocytes) differentiated from PSCs in a closed culture system because each of the individual elements of the instant claims are independently presented by Timmins as embodiments and are taught that they can be combined in various embodiments; therefore a combination of all the elements into a single embodiment would be apparent to an artisan skilled in cell therapy in light of the Supreme Court’s KSR decision (see MPEP 2143 Exemplary Rationale (A)). Regarding the rationale for combining prior art elements according to known methods to yield predictable results, all of the claimed elements were known in the prior art and one skilled in the art could have combined the element as claimed by known methods with no change in their respective functions, and the combination would have yielded predictable results to one of ordinary skill in the art at the time of the invention. Each of the elements mixing the culture medium in the culture vessel hereby allowing the cells to grow in the form of cell aggregates and preventing settling of the cell aggregates and introducing a subsequent culture medium into the culture vessel are taught by Timmins. It would be therefore predictably obvious to use a combination of these elements in said Cardiomyocytes.
With respect to claims 2 and 11, Timmins teaches that the steps g) - i) are repeated or subsequent culture period(s), thereby facilitating parallel serial passaging [0119].
With respect to claim 3, Timmins teaches that the step b) the pluripotent stem cells are introduced in the form of cell aggregates ([0016] of Timmins).
With respect to claims 4 and 6, Timmins teaches that the step b) the amount of pluripotent stem cells is 1 × 106 per ml culture medium, which anticipates stem cells between 1 x 104 - 1 x 106 per ml culture medium [0014]. Wherein at least 20 times more aggregates were processed in a single batch in the STR system ([0182] of Timmins).
With respect to claim 7, Timmins teaches that the cell aggregates collected in step j) have a size of around 150-800 micrometer ([0015] of Timmins).
With respect to claims 8 and 29, Timmins teaches that the wherein the volume of the culture medium in the culture vessel is about 50 mL-2,000 L ([0014] of Timmins).
With respect to claim 9, Timmins teaches that the cell culture comprises about 1x106 cells/mL to 1x1015 cells/mL ([0014] of Timmins).
With respect to claim 12, Timmins teaches that the step c) mixing the culture medium in the culture vessel thereby allowing the cells to grow in the form of cell aggregates at least for 7 days ([0141] of Timmins).
With respect to claim 13, Timmins teaches that the method is performed over a period of 7 days ([0141] of Timmins).
With respect to claim 15, Timmins teaches that in step j) the aggregates collected in the cell reservoir were ([0144] of Timmins). Timmins did not disclose the %volume of collected medium, however Fig. 8 disclose the %recovery of cell aggregates. MEPE 2144.05 states “When there is a design need or market pressure to solve a problem and there are a finite number of identified, predictable solutions, a person of ordinary skill has good reason to pursue the known options within his or her technical grasp. If this leads to the anticipated success, it is likely the product not of innovation but of ordinary skill and common sense” KSR International Co. v. Teleflex Inc., 550 U.S. 398, 82 USPQ2d 1385 (2007). Therefore, it would be obvious to try to collect several %vol culture medium in the culture vessel is collected and a person of ordinary skill in the art to experiment to reach to calculate the collected culture medium in the culture vessel.
With respect to claims 17 and 30, although Timmins discloses is focused on hPSC-derived CMs, the protocol could potentially be adapted to the mass produc-tion of other lineages (i.e., a neuronal cell, a retinal cell, a lung cell, hepatic cells or a pancreatic cell [0060] of Timmins)). The mass production of therapeutically valuable cell types that could benefit from the part of the culture medium that is collected or the culture medium that is collected comprises bank of cells ([0130] of Timmins), wherein the collected cell aggregates could have cell clusters and/or single cells ([0128] of Timmins).
With respect to claim 18, Timmins teaches that the cell inoculated into suspension cultures in mTeSR™l (Stem Cell Technologies) supplemented with Rho-associated protein kinase inhibitor (e.g., Y-27623 (TOCRIS Bioscience)) ([0141] of Timmins).
Regarding claim 19, Timmins does not specifically the culture medium provided in step b) comprises polyvinyl alcohol (PVA). However, such was known in the prior art.
Burridge teaches that a highly efficient method for differentiation of human embryonic stem cells (hESC) and hiPSC to the cardiac lineage (abstract of Burridge). Burridge discloses that polyvinyl alcohol (PVA) synergizes to induce highly efficient cardiac differentiation of hiPSC from 20.663.7% to 68.362.3% (p. 6 left hand Col. 4¶; p. 7 Fig. 4 of Burridge).
MPEP 2143 (A) states that combining prior art elements according to known methods to yield predictable results. The rationale to support a conclusion that the claim would have been obvious is that all the claimed elements were known in the prior art and one skilled in the art could have combined the elements as claimed by known methods with no change in their respective functions, and the combination yielded nothing more than predictable results to one of ordinary skill in the art. KSR, 550 U.S. at 416, 82 USPQ2d at 1395. Accordingly, it would have been obvious to practice the in vitro manufacture method of cardiomyocytes cell of Timmins and include the PVA in the medium as taught by Burridge with a reasonable expectation of success. The POSITA would have been motivated at the time of filing to do so as taught by Burridge because PVA offer efficient and cost-effective universal system for cardiac differentiation of hiPSC allows a potentially unlimited production of functional cardiomyocytes (abstract of Burridge). The POSITA would have had a reasonable expectation of success in combining the teachings of Timmins and Burridge because each of these teachings both successfully differentiates cardiomyocytes from iPSCs. Therefore, the products and method as taught by Timmins et al. in view of Burridge et al. would have been prima facie obvious over the products and method of the instant application. In regard to the reasonable expectation of success in doing so, include the in vitro culture method of Burridge had a reasonable expectation of success since the steps thereof required no more than pipetting the appropriate PVA concentration and cell culture technology.
With respect claim 20, Timmins does not specifically teach the culture media one or more compounds that induce differentiation of the pluripotent stem. However, such was known in the prior art.
Regarding claims 20, 22 and 31, Arauchi teaches a human iPSCs and succeeded in producing over 100 million human cardiomyocytes, which enabled fabrication of functional cardiac tissues in vitro and in vivo and into functional regenerated thyroid follicular cells (TFCs) using scalable suspension culture methods (p. 2 left-hand Col. 2nd ¶). Further, Arauchi discloses that TFCs secreted thyroid hormones and Wnt-pathway activators (i.e., CHIR99021) are inducing differentiation of the PSCs in scalable suspension culture (abstract, p. 3 right-col. 2nd and 3rd ¶ of Arauchi).
MPEP 2143 (A) states that combining prior art elements according to known methods to yield predictable results. The rationale to support a conclusion that the claim would have been obvious is that all the claimed elements were known in the prior art and one skilled in the art could have combined the elements as claimed by known methods with no change in their respective functions, and the combination yielded nothing more than predictable results to one of ordinary skill in the art. KSR, 550 U.S. at 416, 82 USPQ2d at 1395. Accordingly, it would have been obvious to practice the in vitro manufacture method of cardiomyocytes cell of Timmins and include the thyroid hormones and Wnt-pathway activators in the medium as taught by Arauchi with a reasonable expectation of success. The POSITA would have been motivated at the time of filing to do so as taught by Arauchi because differentiation of human iPSCs to functional TFCs that may enable us to fabricate thyroid tissues for regenerative medicine and disease models (abstract of Arauchi). Therefore, it is obvious to utilizing bioreactor will contribute to regenerative medicine development for thyroid dysfunction by allowing effective production of a large enough number of TFCs (p. 9 right-col. 2nd ¶ of Arauchi). The POSITA would have had a reasonable expectation of success in combining the teachings of Timmins and Arauchi because each of these teachings both successfully culture the hPSCs. Therefore, the products and method as taught by Timmins et al. in view of Arauchi et al. would have been prima facie obvious over the products and method of the instant application. In regard to the reasonable expectation of success in doing so, include the in vitro culture method of Arauchi had a reasonable expectation of success since the steps thereof required no more than pipetting the appropriate thyroid hormones concentration and cell culture technology.
Hence, the claimed invention as a whole was prima facie obvious in the absence of evidence to the contrary.
RESPONSE TO ARGUMENTS
Applicant's arguments filed on 16 January 2026 are acknowledged.
Applicant argues that claim rejections - 35 USC§ 102 and 103 over Kempf fails to teach or disclose a "closed culture system" as that term is defined in the present application, understood by one of ordinary skill in the art, and required by amended claim 1 filed on 16 January 2026. See remark pp. 12-14.
Applicant's arguments have been fully considered but they are not persuasive because the new ground of rejection relies on amendment claims and references applied in the prior rejection of record for any teaching or matter specifically challenged in the argument. In the new rejection, Timmins, in view of Burridge, and further in view of Arauchi teaches a method for the in vitro manufacture of a preselected cell type differentiated from a pluripotent stem cell in a closed culture system as recited in the currently amended claims.
Pertinent References
[AltContent: textbox ([img-media_image1.png]
Figure 1. Schematic presentation of the bioreactor set-up used for embryoid body (EB) formation and cardiomyocyte selection.)]The prior art made of record and not relied upon is considered pertinent to applicant's disclosure is the following:
Schroeder et al. (Differentiation and lineage selection of mouse embryonic stem cells in a stirred bench scale bioreactor with automated process control. Biotechnology and bioengineering, 92(7), pp.920-933, 2005) provides embryonic stem (ES) cells can differentiate into functional cardiomyocytes in vitro. ES-derived cardiomyocytes could be used for pharmaceutical and therapeutic applications, provided that they can be generated in sufficient quantity and with sufficient purity (abstract). The genetically engineered ES cells were cultured directly in the stirred bioreactor for 9 days, followed by antibiotic treatment for another 9 days. The protocol resulted in the generation of essentially pure cardiomyocyte cultures, with a total yield of 1.28x109 cells in a single 2 L bioreactor run (abstract; p. 922 and Fig. 1). POSITA would recognize that the Bioreactor in Fig. 1 discloses that it is a closed culture system for preselected-cell type (e.g., cardiomyocytes) differentiated from a pluripotent stem cell (e.g., ES).
Conclusion
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 extension fee 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 date of this final action.
No claims are allowed.
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/MASUDUR RAHMAN/ Patent Examiner, Art Unit 1633
/JEREMY C FLINDERS/ Primary Examiner, Art Unit 1684