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 .
1. The Amendment filed 04/16/2026 has been entered. Claims 34, 35, 37, 38, 41 – 43 and 50 – 57 are pending and under consideration.
Information Disclosure Statement
2. 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.
Objections/Rejections Necessitated by Amendment
Claim Objections
3. Claim 1 is objected to because of the following informalities: in line 5, “Wint” should read “Wnt” for consistency with “Wnt” recited in line 4. Appropriate correction is required.
4. Claim 56 is objected to because of the following informalities: line 1, “are” should read “is” because “aggregate” is singular. Appropriate correction is required.
5. Claim 57 is objected to of the following informalities: line 1, “are” should read “is” because “aggregate” is singular. Appropriate correction is required.
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.
6. Claim 56 and 57 are 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.
7. Claim 56 contains the trademark/trade name Knockout Serum Replacement. Where a trademark or trade name is used in a claim as a limitation to identify or describe a particular material or product, the claim does not comply with the requirements of 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph. See Ex parte Simpson, 218 USPQ 1020 (Bd. App. 1982). The claim scope is uncertain since the trademark or trade name cannot be used properly to identify any particular material or product. A trademark or trade name is used to identify a source of goods, and not the goods themselves. Thus, a trademark or trade name does not identify or describe the goods associated with the trademark or trade name. In the present case, the trademark/trade name is used to identify/describe a serum replacement and, accordingly, the identification/description is indefinite.
8. Claim 57 contains the trademark/trade name Knockout Serum Replacement. Where a trademark or trade name is used in a claim as a limitation to identify or describe a particular material or product, the claim does not comply with the requirements of 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph. See Ex parte Simpson, 218 USPQ 1020 (Bd. App. 1982). The claim scope is uncertain since the trademark or trade name cannot be used properly to identify any particular material or product. A trademark or trade name is used to identify a source of goods, and not the goods themselves. Thus, a trademark or trade name does not identify or describe the goods associated with the trademark or trade name. In the present case, the trademark/trade name is used to identify/describe a serum replacement and, accordingly, the identification/description is indefinite.
9. Claim 57 contains the trademark/trade name Glutamax. Where a trademark or trade name is used in a claim as a limitation to identify or describe a particular material or product, the claim does not comply with the requirements of 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph. See Ex parte Simpson, 218 USPQ 1020 (Bd. App. 1982). The claim scope is uncertain since the trademark or trade name cannot be used properly to identify any particular material or product. A trademark or trade name is used to identify a source of goods, and not the goods themselves. Thus, a trademark or trade name does not identify or describe the goods associated with the trademark or trade name. In the present case, the trademark/trade name is used to identify/describe a culture media additive that is L-alanyl-L-glutamine and, accordingly, the identification/description is indefinite.
Maintained Claim Rejections and New Claim 56 Rejection - 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.
10. Claim(s) 34, 35, 37, 38, 41, 43, 52, 53, 54, and 55 remain rejected and new claim 56 is 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; previously cited), 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; previously cited), hereinafter Lancaster which is cited on the IDS filed 06/02/2022. A transcript of Falan is provided. Although maintained, the rejection is updated in view of the amendment to claims 34 and 43, and new claim 56.
Regarding claim 34 and 35, Falan teaches a method of producing a cerebral organoid comprising culturing aggregates of iPSCs for 18 days (“6 days” and “a further 10-14 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 Rho kinase inhibitor, Wnt signal inhibitor and a TGFβ signal inhibitor for 6 days”, “in the presence of the Wint signal inhibitor and the TGFβ signal inhibitor for a further 10 – 14 days” 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.
PNG
media_image1.png
920
1470
media_image1.png
Greyscale
PNG
media_image2.png
915
1471
media_image2.png
Greyscale
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 Rho kinase inhibitor, Wnt signal inhibitor and a TGFβ signal inhibitor for 6 days”, “in the presence of the Wint signal inhibitor and the TGFβ signal inhibitor for a further 10 – 14 days” 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 “in the presence of a Rho kinase inhibitor, a Wnt signal inhibitor and a TGFβ signal inhibitor for 6 days and in the presence of the Wnt signal inhibitor and the TGFβ signal inhibitor for a further 12 days and then culturing the aggregate for about 17 days without the presence of 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 or the composition of media of claim 56. 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 Rho kinase inhibitor, Wnt signal inhibitor and a TGFβ signal inhibitor for 6 days”, “in the presence of the Wint signal inhibitor and the TGFβ signal inhibitor for a further 10 – 14 days” 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 Y-27632 (“Rho kinase inhibitor”), a Wnt inhibitor and a TGFβ inhibitor from day 0 to day 18 (page 20284, right col. para. 2; Supporting Information page 1, left col. para. 2 – 3; Figure S2A). Kadoshima teaches in Figures S2A that Y-27632 was present during only part of the 18 days while the Wnt and TGFβ inhibitors were present during the entire 18 day period. 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 “in the presence of a Rho kinase inhibitor, a Wnt signal inhibitor and a TGFβ signal inhibitor for 6 days and in the presence of the Wnt signal inhibitor and the TGFβ signal inhibitor for a further 12 days and then culturing the aggregate for about 17 days without the presence of a Wnt signal inhibitor and a TGFβ signal inhibitor” claim 43, Kadoshima teaches after 18 days with Y-27632 (“Rho kinase inhibitor”), 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 1, left col. para. 3; Figure S2A).
Regarding the composition of CDMI of claim 56, Kadoshima teaches the media composition for the first 18 days of culture comprises Glasgow-MEM, 20% KSR (“Knockout Serum Replacement”), nonessential amino acids, pyruvate, 2-mercaptotethanol, penicillin, and streptomycin (Supporting Information page 1, left col. para. 2).
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 Y27632, 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.
11. Claim(s) 42, 50 and 51 remain 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; previously cited), 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; previously cited), 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.
New Claim 57 Rejection - 35 USC § 103
12. Claim(s) 57 is 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; previously cited), 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; previously cited), 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, 55, and 56 above, and further in view of Sakaguchi (Sakaguchi, Hideya, et al. Nature communications 6.1 (2015): 8896), hereinafter Sakaguchi.
Falan in view of Kadoshima and Arlotta make obvious the limitations of claims 34 and 43.
Regarding “CDMI” of claim 57, Kadoshima teaches the media composition for the first 18 days of culture comprises Glasgow-MEM, 20% KSR (“Knockout Serum Replacement”), nonessential amino acids, pyruvate, 2-mercaptotethanol, penicillin, and streptomycin (Supporting Information page 1, left col. para. 2).
Regarding “CDMII” of claim 57, Kadoshima teaches further culturing the aggregates in media containing DMEM/F12, N2, Chemically Defined Lipid Concentrate, Fungizone, penicillin, and streptomycin for 17 days (Supporting Information page 1, left col. para. 3). Kadoshima does not teach “Glutamax” or the concentrations of N2 or Chemically Defined Lipid Concentrate.
Regarding Glutamax and concentrations of N2 and Chemically Defined Lipid Concentrate, Sakaguchi teaches the method of Kadoshima where on day 18 the aggregates were transferred to a non-cell adhesive dish and further cultured in suspension using DMEM/F12, GlutaMAX, 1% N2, 1% Chemically Defined Lipid Concentrate, fungizone, penicillin, and streptomycin for 17 days (page 9, right col. para. 2 – 5; Figure 1a).
Sakaguchi teaches generation of medial pallium from pluripotent stem cells of any species has not yet been achieved (page 2, left col. para. 3). Sakaguchi teaches the method generates dorsomedial telencephalic tissues as well as hippocampal granule- and pyramidal-like neurons (page 2, right col. para. 1). Sakaguchi teaches the hippocampus has been implicated in Alzheimer’s disease and schizophrenia and exploring the molecular mechanism of these disorders has been a challenge (page 9, left col. last para.). Sakaguchi teaches induction of hippocampal neurons from hPSCs should provide a new means to research the molecular biology of hippocampus-relevant disorders (page 9, left col. last para.).
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 Y27632, a Wnt inhibitor and a TGFβ inhibitor to a suspension culture of hESCs aggregates during the first 18 days promoted telencephalic differentiation where the culture media contains Glasgow-MEM, KSR, NEAA, pyruvate, 2-mercaptoethanol, penicillin, and streptomycin followed by culturing the aggregates for 17 days in a culture media containing DMEM/F12, N2, Chemically Defined Lipid Concentrate, Fungizone, penicillin, and streptomycin 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 Sakaguchi regarding culturing day 18 aggregates produced by the method of Kadoshima in media containing DMEM/F12, Glutamax, 1% N2, 1% Chemically Defined Lipid Concentrate, Fungizone, penicillin, and streptomycin, to arrive at the claimed method wherein the aggregate of pluripotent stem cells are cultured for about 18 days in a first medium comprising a Cortical Differentiation Medium I (CDMI), wherein the CDMI comprises Glasgow-MEM, 20% Knockout Serum Replacement, Minimum Essential Medium non-essential amino acids (MEM-NEAA), pyruvate, 2-mercaptoethanol, penicillin, and streptomycin, and further culturing the aggregate for about 17 days in a second medium comprising a Cortical Differentiation Medium II (CDMII) wherein the CDMII comprises DMEM/F12 medium, Glutamax, 1 % N2, 1 % Chemically Defined Lipid Concentrate, fungizone, penicillin, and streptomycin. One would have been motivated to combine the teachings of Falan, Kadoshima, Arlotta, and Sakaguchi in a method of producing dorsal forebrain organoids to study human brain development and neurological disorders as Sakaguchi teaches exploring the molecular mechanism of neurological disorders has been a challenge and Sakaguchi teaches induction of neurons from hPSCs should provide a new means to research the molecular biology of hippocampus-relevant disorders. 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 Sakaguchi teaches the method generates dorsomedial telencephalic tissues as well as hippocampal granule- and pyramidal-like neurons.
Applicant’s Arguments/ Response to Arguments
13. Applicant Argues: Applicant asserts that Falan, Kadoshima, and Arlotta as evidenced by Lancaster whether considered alone or in combination fail to disclose each and every element of the pending claims and Arlotta/Lancaster fail to disclose or suggest culturing the dorsal forebrain aggregates in a spinner flask at about 20% oxygen and 5% CO2 as they do not provide a teaching regarding any defined oxygen value.
Response to Argument: This is not found persuasive because 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 (page 2337, para. 2; Figure 1 and 2f; page 2332, left col. para. 1; page 2331, right col. para. 1) and it is known in the art that standard/ambient air is about 18 to about 20% O2. Thus, absent any teaching by Arlotta/Lancaster that the CO2 incubator regulated O2 concentration to a value outside of standard/ambient air (about 18 to about 20% O2), Arlotta/Lancaster meets the limitation of at about 20% oxygen and 5% CO2 by teaching a tissue culture incubator and 5% CO2 incubator.
Conclusion
No claims 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.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to ZANNA M BEHARRY whose telephone number is (571)270-0411. The examiner can normally be reached Monday - Friday 8:45 am - 5:45 pm.
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, Peter Paras can be reached at (571)272-4517. 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.
/Z.M.B./Examiner, Art Unit 1632
/PETER PARAS JR/Supervisory Patent Examiner, Art Unit 1632
Prosecution Insights