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 .
Continued Examination Under 37 CFR 1.114
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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 11/24/2025 has been entered.
Claim Status
1. The Amendment filed 11/24/2025 has been entered. Claims 34, 35, 37, 38, 41 – 43 and 50 – 55 remain pending and under consideration.
Election/Restrictions
2. Applicant’s election without traverse of Group II (claims 34 – 38 and 41 – 43) in the reply filed on 12/18/2024 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)).
3. Claims 1, 2, 8 – 10, 19 – 21, 30, 32, 33, and 48 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 12/18/2024.
Priority
4. This application is a national stage filing under 35 U.S.C. 371 of International Application No. PCT/US2020/035439, filed 05/29/2020, which claims the benefit of U.S. Provisional Application No. 62/857,802, filed on 06/05/2019, and U.S. Provisional Application Serial No. 62/854,955 filed on 05/30/2019.
Information Disclosure Statement
5. The information disclosure statement (IDS) submitted on 02/11/2026, 11/26/2025, and 11/24/2025 are acknowledged. The submissions are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statements are being considered by the examiner.
6. The listing of references in the specification is not a proper information disclosure statement. 37 CFR 1.98(b) requires a list of all patents, publications, or other information submitted for consideration by the Office, and MPEP § 609.04(a) states, "the list may not be incorporated into the specification but must be submitted in a separate paper." Therefore, unless the references have been cited by the examiner on form PTO-892, they have not been considered.
Withdrawn Claim Objections
7. The objection to claim 34 is withdrawn in view of Applicant’s amendment to the claim.
Withdrawn Claim Rejections
8. The rejection of claims 34 – 35, 38, 41, and 43 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to the claims.
9. The rejection of claims 37 and 42 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to the claims
10. The rejection of claims 34, 37, and 50 – 51 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to the claims
11. The rejection of claims 34 and 52 – 54 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to the claims
12. The rejection of claims 34 and 55 under 35 U.S.C. 103 is withdrawn in view of Applicant’s amendment to the claims.
Rejections Necessitated by Amendment
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.
The factual inquiries 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.
13. Claim(s) 34, 35, 37, 38, 41, 43, 52, 53, 54, and 55 are rejected under 35 U.S.C. 103 as being unpatentable over Falan (Falan. “Falan Courses in Ibro 2015: Cortical circuits – Paola Arlotta”. YouTube, uploaded by Falan, 02/01/2016, https://www.youtube.com/watch?v=GrR89E2jMcs), hereinafter Falan in view of Kadoshima (Kadoshima, Taisuke, et al. Proceedings of the National Academy of Sciences 110.50 (2013): 20284-20289; previously cited), hereinafter Kadoshima in view of Arlotta (WO-2017117547-A1; previously cited), hereinafter Arlotta as evidenced by Lancaster (Lancaster, Madeline A., et. al. Nature protocols 9.10 (2014): 2329-2340.), hereinafter Lancaster which is cited on the IDS filed 06/02/2022. A transcript of Falan is provided.
Regarding claim 34 and 35, Falan teaches a method of producing a cerebral organoid comprising culturing aggregates of iPSCs for 18 days (“culturing an aggregate of pluripotent stem cells” and “16-20” days of claim 34; “about 18 days” of claim 35) followed by culturing in a spinner flask for more than 35 days (“culturing the dorsal forebrain aggregate in a spinner flask” and “for about 35 days more” of claim 34) (screenshots below; page 47 – 50 of the transcript at timestamp 31:52 – 34:05). Falan does not teach “in suspension in the presence of a Wnt signal inhibitor and a TGFβ signal inhibitor” or “at about 20% oxygen and 5% CO2” of claim 34.
Regarding claim 37, Falan teaches the method produces organoids that are positive for CTIP2, SATB2, TLE4, and BRN2 (“corticofugal projection neurons”, “callosal projection neurons”, “immature corticofugal projection neurons”, “immature callosal projection neurons”, “immature projection neurons”) (second screenshot below; page 49 of the transcript timestamp 33:18) but does not teach the remainder of the cells recited in the claim.
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Regarding claim 54 and 55, Falan teaches the method starts with iPSCs taken from anyone (“human” of claim 54) and they can be obtained from directed reprogramming from skin and organoids from patients with specific diseases of from stem cells that are genetically modified to carry a mutation associated with the disease (claim 55) (page 47 of the transcript, timestamp 31:52 – 32:10 and 33:54 – 33:59).
Falan does not teach “in suspension in the presence of a Wnt signal inhibitor and a TGFβ signal inhibitor” or “at about 20% oxygen and 5% CO2” of claim 34 or “cycling progenitors”, “intermediate progenitor cells, “outer radial glia”, “Cajal-Retzius neurons” or “radial glia” of claim 37 or “162 days or more” of claim 38 or “cycling progenitors” of claim 41 or culturing without a Wnt signal inhibitor and a TGFβ signal inhibitor of claim 43 or “about 9 months or more” of claim 52 or “about 1 year or more” of claim 53. However, Falan teaches the method produces organoids with neuroectoderm similar to that seen during normal human brain development (page 47 – 48 of the transcript; timestamp 32:10 – 32:30). Falan teaches the organoids are inside spinning bioreactors in an incubator and you spin them for months and months and months and they grow and continue to develop (page 48 of the transcript; timestamp 32:35 – 32:41). Falan teaches the organoids have regions that are cortical plate-like, VZ-like, layer V-like, LV, layer VI-like, upper layers-like, and deep layers-like (second screenshot above).
Regarding “in suspension in the presence of a Wnt signal inhibitor and a TGFβ signal inhibitor” of claim 34, Kadoshima teaches a method of producing brain organoids comprising a first step of culturing aggregates of hESCs in low-cell-adhesion plates in suspension culture with a Wnt inhibitor and a TGFβ inhibitor from day 0 to day 18 (page 20284, right col. para. 2; Supplemental Information page 4, left col. para. 1; Figure S2A). Kadoshima teaches the addition of a Wnt inhibitor and a TGFβ inhibitor during the first 18 days promoted telencephalic differentiation (page 20284, right col. para. 2). Kadoshima teaches that the method improves on previous methods to produce smooth organoids at day 7 in Figure S2A’. Kadoshima does not teach “at about 20% oxygen and 5% CO2”.
Regarding claim 37, Kadoshima teaches the method produces organoids containing intermediate progenitors, outer radial glia, radial glia, and Cajal-Retzius neurons (Figure 3, 5, and S5; page 20286, right col. para. 2 – 3; page 20289, left col. para. 3; page 20286, right col. para. 1; Abstract). Kadoshima teaches the method allows for self-formation of multilayered neuroepithelium that includes subplate, cortical plate, and Cajal-Retzius cell zones and ventricular, subventricular and intermediate progenitor zones in the same apical-basal order as seen in the human fetal cortex in the early second trimester (Abstract). Kadoshima does not teach “cycling progenitors”.
Regarding claim 43, Kadoshima teaches after 18 days with a Wnt inhibitor and a TGFβ inhibitor, the aggregates are cultured for up to 70 days without a Wnt inhibitor and a TGFβ inhibitor (Supplemental Information page 7, left col. para. 3).
Kadoshima does not teach “at about 20% oxygen and 5% CO2” of claim 34 or “cycling progenitors” of claim 37 and 41 or “162 days or more” of claim 38 or “about 9 months or more” of claim 52 or “about 1 year or more” of claim 53. However, Kadoshima teaches a detailed understanding of early human corticogenesis remains elusive because of the limited access to fetal brain tissues (page 20284, left col. para. 2). Kadoshima teaches the method may be applicable to studies of the inside-out pattern formation in the human fetal cortex including the pathogenesis of lissencephaly (page 20289, left col. para. 1). Kadoshima teaches the 3D culture allows highly selective induction and long-term growth of hESC-derived cortical neuroepithelium that self-organizes (Abstract). One would have been motivated to combine the teachings of Falan and Kadoshima because both teach methods of producing dorsal forebrain organoids that can be used for studying brain development and disease.
Regarding “at about 20% oxygen and 5% CO2” of claim 34 and claims 38, 52, and 53, Arlotta teaches a method of producing brain organoids from human pluripotent stem cells comprising following neural induction embedding organoids in Matrigel and culturing the organoids in a spinner flask for up to 13 months (claims 38, 52, 53) where the rotation of the spinner flask is a rate that allows sufficient oxygen diffusion in the medium and at the same time preserves the integrity of the brain organoids (page 69, last line; page 70, lines 1 – 6; Figure 1A; page 46, lines 2 – 8). Arlotta teaches the formation of embryoid bodies and organoids with smooth edges (page 46, lines 26 – 27; Figure 1A, 3E, and 5A). Arlotta teaches the method was a modification of Lancaster (page 59, lines 7 – 17; page 69, lines 25 – 26) where the organoids were cultured in a spinning bioreactor installed in a CO2 incubator with 5% CO2 as evidenced by Lancaster (“at about 20% oxygen and 5% CO2” of claim 34) (page 2337, para. 2; Figure 1 and 2f; page 2332, left col. para. 1; page 2331, right col. para. 1).
Regarding “cycling progenitors” of claim 37 and claim 41, Arlotta teaches the organoids show expression of dorsal forebrain markers and show expression markers for glial populations over 1 to 9 months of culture which includes PAX6 that is a marker for radial glial cells (page 8, lines 24 – 26; Figure 5B – 5C; page 41, line 22). Arlotta teaches the organoids comprise cells which express Reelin that is expressed in Cajal-Retzius cells (page 41, lines 31 – 33). Arlotta teaches the organoids comprise cells which express MKI67 where MKI67 and TOP2A act as markers of highly proliferative progenitors (“cycling progenitors” of claim 37 and 41) (page 43, lines 33 – 34). Arlotta teaches the organoids at 6 months expressed cortical projection neuron markers including markers of corticofugal and callosal projection neurons (Figure 5C; page 59, last line; page 60, lines 1 – 4). Arlotta teaches the data regarding the types of neurons in the organoids indicate a developmental sequence of cell types and regional identities that broadly follows the temporal progression observed in vivo (page 60, lines 2 – 4).
Arlotta teaches human brain development and neurodevelopmental disorders are poorly understood processes and studies using human and primate brain tissue have been limited by practical and ethical concerns related to tissue availability, expansion, and manipulation (page 1, lines 1 – 7). Arlotta teaches brain organoids offer an opportunity to study both normal brain development and complex human diseases that affect multiple cell types, the interactions between them, and the function of neuronal circuits (page 1, lines 8 – 13).
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 Falan regarding a method of producing cerebral organoids by culturing iPSC aggregates for 18 days followed by culturing for more than 35 days in a spinner flask with the teachings of Kadoshima regarding a method of producing brain organoids comprising the addition of a Wnt inhibitor and a TGFβ inhibitor to a suspension culture of hESCs aggregates during the first 18 days promoted telencephalic differentiation with the teachings of Arlotta regarding a method of producing brain organoids from human pluripotent stem cells comprising culturing organoids in a spinner flask in an incubator for up to 13 months to arrive at the claimed method of producing a reproducible dorsal forebrain organoid, comprising:
a first step consisting essentially of culturing an aggregate of pluripotent stem cells in suspension in the presence of a Wnt signal inhibitor and a TGFβ signal inhibitor for 16-20 days in a low attachment plate, thereby forming a dorsal forebrain aggregate; and
a second step comprising culturing the dorsal forebrain aggregate in a spinner flask at about 20% oxygen and 5 % CO2 for about 35 days or more, thereby forming a reproducible dorsal forebrain organoid. One would have been motivated to combine the teachings of Falan, Kadoshima, and Arlotta in a method of producing dorsal forebrain organoids to study human brain development and neurodevelopmental disorders as Kadoshima teaches a detailed understanding of early human corticogenesis remains elusive because of the limited access to fetal brain tissues and Arlotta teaches human brain development and neurodevelopmental disorders are poorly understood processes and studies using human and primate brain tissue have been limited by practical and ethical concerns related to tissue availability, expansion, and manipulation. One would have a reasonable expectation of success in combining the teachings as Falan teaches the method produces organoids with neuroectoderm similar to that seen during normal human brain development and Falan teaches organoids can be produced with iPSCs from patients with specific diseases of from stem cells that are genetically modified to carry a mutation associated with the disease and both Kadoshima and Arlotta teach the method produces brain organoids with neurons found in the dorsal forebrain and Kadoshima teaches the method allows for highly selective induction and long-term growth of hESC-derived cortical neuroepithelium that self-organizes.
14. Claim(s) 42, 50 and 51 is/are rejected under 35 U.S.C. 103 as being unpatentable over Falan (Falan. “Falan Courses in Ibro 2015: Cortical circuits – Paola Arlotta”. YouTube, uploaded by Falan, 02/01/2016, https://www.youtube.com/watch?v=GrR89E2jMcs), hereinafter Falan in view of Kadoshima (Kadoshima, Taisuke, et al. Proceedings of the National Academy of Sciences 110.50 (2013): 20284-20289; previously cited), hereinafter Kadoshima in view of Arlotta (WO-2017117547-A1; previously cited), hereinafter Arlotta as evidenced by Lancaster (Lancaster, Madeline A., et. al. Nature protocols 9.10 (2014): 2329-2340.), hereinafter Lancaster which is cited on the IDS filed 06/02/2022 as applied to claims 34, 35, 37, 38, 41, 43, 52, 53, 54, and 55 above, and further in view of Quadrato (Quadrato, Giorgia, et al. Nature 545.7652 (2017): 48-53; previously cited), hereinafter Quadrato which is cited on the IDS filed 06/02/2022.
Falan in view of Kadoshima and Arlotta make obvious the limitations of claims 34, 37, and 41.
Regarding claim 42, Arlotta teaches the addition of BDNF to the final differentiation medium allows the culture period to be prolonged, thereby limiting premature cell death and enabling long-term, progressive development for 9 – 13 months (page 46, lines 19 – 21). However, Arlotta does not teach hypoxia or apoptosis levels of the cells.
Regarding claim 50, Kadoshima does not teach the presence of astroglia or cycling interneurons and Arlotta teaches astroglia became apparent only at 3 months (page 59, lines 30 – 31). However, Kadoshima does not teach percentages of cells.
Regarding “apoptotic or hypoxic cells” of claim 42, Quadrato teaches a method of forming brain organoids comprising neural induction of hiPSCs followed by embedded embryoid bodies in Matrigel and transferring to spinning bioreactors for long-term culture up to 13 months and the organoids do not become hypoxic and levels of programmed cell death remain relatively low up to nine months (page 48, right col. para. 2; Extended data Figure 1a; Figure 1a and Figure legend; page 54, left col. para. 2 – 3).
Regarding claim 50, Quadrato teaches the distribution of the different types of cells present in the 6 month old organoids in Figure 1b, c and the percentage of the different types of cells in Extended Data Figure 5 where Org3B contains 15% corticofugal projection neurons, Org3A contains 37% callosal projection neurons, and Org3A contains 7% intermediate progenitor cells.
Regarding claim 51, Quadrato teaches 6 month old organoid Org3B contains 23% callosal projection neurons and Org3A contains 7% intermediate progenitor cells (Extended Data Figure 5).
Quadrato teaches in vitro models of the developing brain such as 3D brain organoids offer an unprecedented opportunity to study aspects of human brain development and disease (Abstract). Quadrato teaches the analyses of cell types in the organoids indicate that distinct cell types generated in the organoids transcriptionally resemble the appropriate endogenous counterparts from the human fetal cortex (page 50, left col. para. 2). Quadrato teaches human brain organoids have enormous potential to serve as in vitro models of the human brain (page 52, left col. last para.). Quadrato teaches that the 3D brain organoids have the potential to model higher-order functions of the human brain, such as cellular interactions and neural circuit dysfunctions related to neurodevelopmental and neuropsychiatric pathologies (page 53, left col. last para.). Quadrato teaches it is essential to understand whether 3D brain organoids can continue to develop in culture past early development events to allow not only the generation of endogenous cellular diversity, but also the maturation of neuronal networks, which will be needed to apply brain organoids to studies of late developmental events, such as complex cellular interactions and, most notably higher-order brain functions that rely on functional neural networks (page 48, left col. para. 4).
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 Falan regarding a method of producing cerebral organoids by culturing iPSC aggregates for 18 days followed by culturing for more than 35 days in a spinner flask with the teachings of Kadoshima regarding a method of producing brain organoids comprising the addition of a Wnt inhibitor and a TGFβ inhibitor to a suspension culture of hESCs aggregates during the first 18 days promoted telencephalic differentiation with the teachings of Arlotta regarding a method of producing brain organoids from human pluripotent stem cells comprising culturing organoids in a spinner flask in an incubator for up to 13 months with the teachings of Quadrato regarding brain organoids cultured in a spinner flask for 6 months comprise 7% intermediate progenitor cells and 23% callosal projection neurons to arrive at the claimed method where dorsal forebrain organoids comprise 7% intermediate progenitor cells and where dorsal forebrain organoids comprise 23% callosal projection neurons at 6 months. One would have been motivated to combine the teachings of Falan, Kadoshima, Arlotta, and Quadrato in a method of producing dorsal forebrain organoids to study human brain development and neurodevelopmental disorders as Quadrato teaches human brain organoids have enormous potential to serve as in vitro models of the human brain. One would have a reasonable expectation of success in combining the teachings as Falan teaches the method produces organoids with neuroectoderm similar to that seen during normal human brain development and Falan teaches organoids can be produced with iPSCs from patients with specific diseases of from stem cells that are genetically modified to carry a mutation associated with the disease and Quadrato teaches the analyses of cell types in the organoids indicate that distinct cell types generated in the organoids transcriptionally resemble the appropriate endogenous counterparts from the human fetal cortex.
Applicant’s Arguments/ Response to Arguments
15. Applicant Argues: On page 5, last para. and page 6, Applicant asserts that Eguchi does not teach amended claim 34 because Eguchi requires FGF2 and the organoids of Eguchi do not have uniformity in size or shape.
Response to Argument: The previous rejection of the claims using the teachings of Eguchi have been withdrawn and new rejections are set forth above. In the new rejection, Falan in view of Kadoshima make obvious the method of claim 34. It is noted that claim 34 has been amended to recite “a reproducible dorsal forebrain organoid”, however, “reproducible” does not add structure to the dorsal forebrain organoid. The organoids shown in the screenshots are consistent in size and shape and the method of Falan comprises a spinner flask required in amended claim 34 to form a reproducible dorsal forebrain organoid.
Applicant Argues: On page 7, paragraph 2, Applicant asserts one of skill in the art would not have sought to combine Arlotta with Eguchi because the methods of Eguchi were specifically formulated not to utilize special equipment such as a spinner flask.
Response to Argument: The previous rejection of the claims using the teachings of Eguchi have been withdrawn and new rejections are set forth above. One of skill in the art would have sought to combine Falan with Arlotta because both teach methods of producing brain organoids using spinner flasks in an incubator.
Applicant Argues: On page 7, last para., and page 8, para. 2 and last para., Applicant asserts Kadoshima, Quadrato, or Mariani do not teach amended claim 34.
Response to Argument: In the new rejections set forth above, Falan in view of Kadoshima make obvious the method producing dorsal forebrain organoids by culturing an aggregate of pluripotent stem cells in suspension in the presence of a Wnt signal inhibitor and a TGFβ signal inhibitor for 18 days in a low attachment plate, thereby forming a dorsal forebrain progenitor aggregate, and culturing the dorsal forebrain progenitor marker-positive aggregate in a spinner flask at about 20% oxygen and 5% CO2 in a tissue culture incubator for about 35 days or more. Falan teaches organoids at 60 and 105 days are positive for CTIP2, a dorsal forebrain marker (second screenshot). Quadrato teaches a method of producing brain organoids using a spinner flask for prolonged culture and the brain organoids at 3 and 6 months are positive for cells of the dorsal forebrain. The previous rejection using the teachings of Mariani has been withdrawn.
Conclusion
No claims allowed.
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/ZANNA MARIA BEHARRY/Examiner, Art Unit 1632