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 .
Status of the Claims
Claims 12, 22, and 26 are currently pending.
Claims 12 and 22 are amended.
Claims 1-11, 13-21, and 23-25 are cancelled.
Claims 12, 22, and 26 have been considered on the merits.
Withdrawn Rejections
The 112(b) rejection made onto claims 1 and 4 is withdrawn in light of the cancellation of claims 1 and 4 in the reply filled on 12/02/2025.
New Rejections Necessitated by Amendment
Claim Objections
Claim 26 is objected to because of the following informalities: “then seeding the mixture” in line 13 can be amended to “seeding the mixture” to maintain the same structure of active step as the “providing”, “coating”, “mixing”, and “incubating” steps of the method. Appropriate correction is required.
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.
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 12, 22, and 26 are rejected under 35 U.S.C. 103 as being unpatentable over Mangan et al (US 20160116459 A1) in view of Aiba et al (WO 2019098256, WIPO English translation), both of which are references of record.
Regarding claim 26, Mangan teaches providing a cell culture container containing an electrode array arranged on the culture surface and coating the culture surface with at least one coating agent such that an adhesion area is obtained, the coating agent being polyethyleneimine (PEI) ([0080]-[0085]). Mangan teaches mixing neurons and astrocytes to form a co-culture, wherein the percentages of neurons and astrocytes in the mixture can range anywhere from 10-90% neurons and anywhere from 1-55% astrocytes and any range derivable therein, which meets the claimed limitation of anywhere from 66.7-80% pluripotent stem cells/neurons and 20-33.3% astrocytes (ratios of pluripotent stem cells or neurons : numbers of astrocytes of 2:1 to 4:1) ([0007]/[0034]). Mangan teaches that “when iPS cells are “primed” in the absence of TeSR growth factors, i.e., cultured in any medium that does not have basic fibroblast growth factor (bFGF) and transforming growth factor β (TGFβ), for several days prior to aggregate formation, the cells can develop into the neural lineage with purity, rapidity and consistency” which meets the limitation of “mixing treated pluripotent stem cells subjected to neuronal differentiation treatment before differentiation into neurons”. Mangan teaches that “cells that are undifferentiated, pluripotent, multipotent, or neural progenitor cells may be placed in a well comprising a multielectrode array and then differentiated into neural cells” which meets the limitations of and “wherein said treated pluripotent stem cells and said astrocytes are the only cells present in the mixture” ([0042]). Mangan teaches seeding the mixture in the electrode array culture surface ([0120]). Mangan teaches incubating the cell culture container such that the mixture of cells are made to adhere to the culture surface ([0110]), which also meets the limitation of adhering to the culture surface “only in the adhesion area” because the “adhesion area” is described as the coated culture surface in lines 4-5 of claim 1, and Mangan teaches coating an electrode array 48 well plate and then culturing on said plate ([0079]-[0110]). Additionally, Mangan repeats throughout the procedure to not add medium to the plate too quickly or it “may dislodge the adhered neurons” ([0079]-[0110]). Mangan teaches incubation periods of up to 12 days ([0128]).
Regarding the limitation “wherein the adhesion area is 3.14 mm2 to about 28.2 mm2 per 80,000 total cells” in claim 26, the instant specification discloses that the adhesion area is an adhesion area of the cells to the culture surface in which an aggregation of cell bodies is present ([0045]) and further discloses that the adhesion area of the cells decreases as the aggregation level of the cells increases ([0055]). Thus, the adhesion area is interpreted according to the instant specification as a variable that is not a defined structural component of the cell culture container, but a value that is only obtained when the cells adhere to the culture surface. Therefore, the adhesion area is interpreted as a result of practicing the claimed method and could have been determined and modified by a person of ordinary skill in the art by adjusting the culture surface or the culture medium to cause an increase or decrease in the aggregation of the cell.
Regarding the limitation “wherein the adhesion of said neurons to the cell culture container is improved by the mixing “ and “wherein when said treated pluripotent stem cells and said astrocytes, instead of initial differentiated neurons and said astrocytes, are subjected to the mixing and the seeding, separation of said neurons from the culture surface is inhibited compared to the mixing and seeding of said initial differentiated neurons and said astrocytes” of claim 26, Mangan teaches the method step of mixing the treated pluripotent stem cells and astrocytes requiring that the pluripotent stem cells be included in the mixture prior to seeding and differentiation into neurons. The claimed wherein clauses recite the intended result of the method step of mixing the treated pluripotent stem cells and astrocytes prior to seeding rather than requiring an additional step be performed. MPEP 2111.04 states “Claim scope is not limited by claim language that suggests or makes optional but does not require steps to be performed” and that a such a clause ‘"in a method claim is not given weight when it simply expresses the intended result of a process step positively recited.” Therefore since these claims only recite the results of the steps, then art reading on claim 26, specifically the step of mixing, will also read on these results since performing the same steps will inherently lead to the same results in the absence of evidence to the contrary including unexpected results.
Regarding claim 12, Mangan teaches that the treated pluripotent stem cells are iPS cells ([0042]-[0043]).
Regarding claim 22, Mangan teaches wherein the cell-culture container is a microelectrode array (MEA) plate ([0038]).
Mangan fails to disclose a total cell number of the pluripotent stem cells and the astrocytes per unit area of the culture surface of 2,500 to 300,000 cells/cm2, as recited in instant claim 26.
However, Aiba teaches a nerve cell device (cell-containing container) for culturing pluripotent stem cell derived-nerve cells (neurons) on a cell scaffold ([0007]) and a method of making and using the nerve cell device for improving cell adhesion and reducing cell separation from a MEA ([0009]). Furthermore, Aiba teaches that neurons can be cultured with astrocytes ([0017]) and, in example 9, Aiba teaches seeding nerve cells at a density of 1.0 x 105 (100,000) cells/cm2 and 3.0 x 105 (300,000) cells/cm2, which falls within the range of total cell number per unit area of the culture surface disclosed in instant claim 26 ([0031]). While Aiba teaches PSC-derived nerve cells ([0008]/[0017]), Aiba does not teach incubating pluripotent stem cells destined to differentiate into neurons with astrocytes.
Nonetheless, it would have been obvious to a person of ordinary skill in the art at the effective filing date of the claimed invention to seed a total cell number of the pluripotent stem cells and the astrocytes per unit area of the culture surface of 2,500 to 300,000 cells/cm2 as recited in instant claim 26, because Aiba teaches a method based on nerve cells, similar to Mangan’s method, which discloses cell seeding densities that could be applied in experimental designs where PSCs will be differentiated to nerve cells at a certain step of the experiment in co-culture with astrocytes. One of ordinary skill in the art would have been motivated to combine Mangan’s teachings in combination with Aiba’s because a cell culture using nerve cells and astrocytes on a MEA, or PSCs and astrocytes on a MEA, such as in Aiba’s disclosure and Mangan’s respectively, would serve as a reasonable example for culturing PSCs mixed with astrocytes on a MEA for the same purpose of obtaining a neuron population in a cell culture container.
Moreover, the instantly recited values would be within the realm of routine experimentation. Generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature is critical. "[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation." In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955). See MPEP § 2144.05 part II A. It would have been obvious to one of ordinary skill in the art at the time Applicants' invention was made to determine all operable and optimal total cell number because cell seeding density is an art-recognized, result-effective variable known to affect the development of a cell culture, which would have been optimized in the art to obtain the desired results.
Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the time the invention was made, especially in the absence of evidence to the contrary.
Response to Arguments
Applicant's arguments filed 12/02/2025 have been fully considered but they are not persuasive.
Applicant argues (Remarks, pg. 8) that Mangan does not disclose the culture of iPS cells with astrocytes and not neurons and instead discloses that neurons can be cultured with iPS cells in [0009].
In response, the argument is not found persuasive because although Mangan teaches that neurons can be incubated with IPS cells in an embodiment of the invention this is not required and is presented as an alternative embodiment ([0009]). Additionally, Mangan teaches that “A variety of methods are available for generating neurons and/or astrocytes for use in various aspects of the present invention. In some embodiments, neurons (e.g., GABAergic, glutamatergic, or dopaminergic neurons, etc.) or astrocytes may be cultured produced from pluripotent cells such as iPS cells or stem cells. In some embodiments, neurons may be differentiated prior to culture in a well comprising a multielectrode array (e.g., a MEA); in other embodiments, cells that are undifferentiated, pluripotent, multipotent, or neural progenitor cells may be placed in a well comprising a multielectrode array and then differentiated into neural cells.”([0042]). The underlined portion of this excerpt from Mangan meets the limitations of and “mixing treated pluripotent stem cells subjected to neuronal differentiation treatment before differentiation into neurons” and “wherein said treated pluripotent stem cells and said astrocytes are the only cells present in the mixture” ([0042]). The underlined portion also exemplifies that Mangan is not only teaching the addition of iPS cells in the presence of previously differentiated neurons as implied by Applicant’s argument. Therefore, the argument is not found persuasive.
Applicant argues (Remarks, pg. 9) that the cited reference does not disclose the claim limitation of an adhesion area in which said neurons and said astrocytes adhere to the culture surface is 3.14-28.2 mm2 per 80,000 total cells including said neurons and said astrocytes. Applicant argues that the specification describes that when PSCs are induced to differentiate within the culture apparatus it is possible to further inhibit separation of neurons. Applicant concludes that the stable detection of action potentials within the range of adhesion area claimed is difficult to predict and is unexpected, “furthermore, by seeding incompletely differentiated pluripotent cells and astrocytes, preventing detachment unexpectedly occurs”.
In response, the argument is not found persuasive. With regards to the claim limitation “wherein the adhesion area of the neurons and the astrocytes to the culture surface is 3.14 mm2 to about 28.2 mm2 per 80,000 total cells” in claim 26, the instant specification discloses that the adhesion area is an adhesion area of the cells to the culture surface in which an aggregation of cell bodies is present ([0045]) and further discloses that the adhesion area of the cells decreases as the aggregation level of the cells increases ([0055]). Thus, the adhesion area is interpreted according to the instant specification as a variable that is not a defined structural component of the cell culture container, but a value that is only obtained when the cells adhere to the culture surface. Therefore, the adhesion area is interpreted as a result of practicing the claimed method and could have been determined and modified by a person of ordinary skill in the art by adjusting the culture surface or the culture medium to cause an increase or decrease in the aggregation of the cell.
In response to applicant's argument that the references fail to show certain features of the invention, it is noted that the features upon which applicant relies (i.e., stable detection of action potentials) are not recited in the rejected claim(s). Although the claims are interpreted in light of the specification, limitations from the specification are not read into the claims. See In re Van Geuns, 988 F.2d 1181, 26 USPQ2d 1057 (Fed. Cir. 1993).
Additionally, Applicant argues that the prevention of detachment is an unexpected result. This is not found persuasive. Mangan explicitly teaches the differentiation of PSCs within the culture container and therefore, the method of Mangan would inherently result in the increase in adherence seen.
Conclusion
No claims are 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.
Examiner Contact Information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to CONSTANTINA E STAVROU whose telephone number is (571)272-9899. The examiner can normally be reached M-F 8:00-5:00.
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CONSTANTINA E. STAVROU
Examiner
Art Unit 1632
/ANOOP K SINGH/Primary Examiner, Art Unit 1632