CTNF 18/559,119 CTNF 99262 Notice of Pre-AIA or AIA Status 07-03-aia AIA 15-10-aia The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA. 12-151 AIA 26-51 12-51 Status of Claims Claims 1-38 are pending. Priority Claims 1-38 are a 371 of PCT/US 22/28385 filed on May 9, 2022, which has priority to PRO 63/185,961 filed on May 7, 2021. Information Disclosure Statement The information disclosure statement(s) (IDS) submitted on November 6, 2023, was filed before the mailing of the First Office Action on March 28, 2026. The Non-Patent Literature is in compliance with the provisions of 37 CFR 1.97 and are being considered by the examiner. Claim Objections Claim 26 is objected for the following reasons: Claim 26 recites “negative for expression Oct4” should read “negative for the expression of Oct4”. Claim 26 recites “and negative for expression Nanog” should read “and negative for the expression of Nanog”. Examiner’s note: While Applicant’s use of “the cells” provides sufficient antecedent basis for claims 6-10, 15, 17, 18, 20-22, Applicant is advised that replacing “the cells” with “ENCCs” would improve clarity and avoid potential confusion, particularly because the base claim, e.g. claim 1, refers both to enteric neural crest cells and the stem cells from which they are derived from. The same rational is applied to claims 35 and 36 given that claim 27 refers to both enteric neural crest cells and stem cells. Claim Rejections - 35 USC § 112 07-30-01 AIA The following is a quotation of the first paragraph of 35 U.S.C. 112(a): (a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention. The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112: The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention. 07-31-02 AIA Claim 22 is rejected under 35 U.S.C. §112(a) or 35 U.S.C. §112 (pre-AIA), first paragraph, as failing to comply with the enablement requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention. Nature of the Invention: Claim 22 is directed to a method of treating enteric neuropathy using stem cell-derived enteric neural crest cells where “the cells” are genetically modified to “evade immune system surveillance”. Breadth of Claims: Claim 22 broadly recites “the cells are genetically modified to evade immune system surveillance” without limitation to the specific genes modified, the type or extent of modification, e.g. knock-out, knock-in, overexpression, or a combination of some sort, which immune pathways are targeted, and whether or how the modifications are retained after differentiation into enteric neural crest cells. Amount of Direction or Guidance Present in the Specification: The specification provides minimal guidance regarding immune system evasion. The specification specifically states “the cells have been genetically modified to evade immune system surveillance” [pg. 2, Line 29]. In the same paragraph, the specification generically states that stem cells my be genetically modified, but does not describe how such modification achieves immune evasion. The only genes identified are on pg. 25, Line 15, including the tables, include GDNF, integrin, FAK, ITGB4, RET, and/or EDNRB associated with cell survival, migration, and neural development. There is no teaching where human leukocyte antigen pathways are modified, modifications that allow antigen presentation suppression, or modulating immune checkpoint pathways. Presence or Absence of Working Examples: The specification contains no working examples demonstrating genetic modification of stem cells that are capable of evading immune surveillance, no examples of either in vitro or in vivo where successful evasion was achieved, or retention of immune-evasive properties after differentiation into enteric neural crest cells. Relative Skill in the Art: It is argued that the relative skill in the art, is that of a scientist with several years’ experience in the field, but that the Art itself is a recognition of what is understood by the Artisan, and thus, as seen below, does not make the breadth of the claims more predictable. The Predictability or lack thereof in the art: The art is highly unpredictable. As explained by Elmusa et al., immune rejection remains a fundamental barrier due to human leukocyte antigen (HLA) mismatches [Abstract, Engineering the ‘one-for-all’ cell: Molecular approaches to universal hypoimmunogenic pluripotent stem cells, Biomedicine Advances , 2025]. Elmusa et al. further states that immune recognition occurs through multiple pathways, including direct and indirect recognition, and that suppression of human leukocyte antigen class I expression triggers natural killer cell “missing self” responses [pg. 184, left column]. Elmusa et al. further states that even partial strategies that involve HLA-A/B knockout with HLA-C retention fail to eliminate CD8+T cell responses and may still permit CD4+ T cell activation via HLA class II [pg. 187, right column]. This demonstrates that even the current art recognizes that accomplishing a cell that is capable of immune system evasion remains difficult. Furthermore, the art does not establish that such modifications would be retained after differentiation into specialized cell types such as enteric neural crest cells. This further complicates the unpredictability. Quantity of Experimentation Needed: For a person of ordinary skill to practice the full scope of the invention, the artisan would need to identify appropriate immune evasion targets, e.g. HLA pathways, checkpoint molecules, design and implement multiple genetic modifications, evaluate immune interactions across T cell, NK cells, and antigen presenting cells, differentiate modified iPSCs into enteric neural crest cells, and determine whether the immune-evasive properties are retained post-differentiation. Conclusion: Given this, the specification fails to provide sufficient disclosure to allow one of ordinary skill in the art to make and use the claimed invention without undue experimentation . 07-31-01 Claim 22 is rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention. Claim 22 uses the generic phrase “cells are genetically modified to evade immune system surveillance” as it relates to induced pluripotent stem cells being differentiated into enteric neural crest cells that are then administered to treat enteric neuropathy. The specification provides antecedent basis for “cells are genetically modified to evade immune system surveillance” [pg. 2, line 29]. Applicant’s specification states “in some embodiments, the cells have been genetically modified to evade immune system surveillance”. Additionally, the specification in the same paragraph states that cells can be genetically modified to overexpress and/or under express a gene or gene product of interest. There is no other mention in the specification related to stem cells or enteric neural crest cells being modified in such a way that would allow these cells to evade “immune system surveillance”. The only example provided in the specification of genetic modifications is related to cell survival, migration, and neural development [pg. 25, line 15]. Furthermore, current art consistently discusses the many obstacles that remain in creating cells that are capable of evading immune system surveillance, whether it be related to direct immune responses or indirect immune responses. For example, Elmusa et al., discussing several methods to explore and the remaining challenges of hypoimmunogenic stem cells, discloses that immune rejection following allogenic transplantation remains a fundamental barrier due to mismatches in human leukocyte antigen (HLA) mismatches [Abstract, Engineering the ‘one-for-all’ cell: molecular approaches to universal hypoimmunogenic pluripotent stem cells, Biomedicine Advances , 2025]. Emulsa et al. goes on to discuss the obstacles relating to such things as knocking out expression of one characteristic typically results in an immune response related to “missing self” responses from other cells [Pg. 184, left column]. Emulsa et al. further details that there are still fundamental obstacles that need to be overcome before immune evasion can be achieved, including sophisticated genetic engineering [Conclusion]. Given this and the lack of direction or examples provided in the specification, a person of ordinary skill would not understand Applicant to be in possession of any cell capable of evading immune system surveillance. Given the generic scope of “cells are genetically modified to evade immune system surveillance” as it relates to the differentiation of stem cells into neural crest cells capable of evading immune system surveillance and the absence of any teachings, the Artisan would not understand Applicant to be in possession of the generic scope of “cells are genetically modified to evade immune system surveillance”. 07-31-01 Claim 1, 6-18, 20-22, 23, 27, and 32-38 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention. Claim 1 uses the generic term “stem cell” as it relates to differentiating induced pluripotent stem cells into enteric neural crest cells for administration in subjects with enteric neuropathy. The same generic scope for “stem cell” is present in each of the dependent claims, i.e. 6-18, and 20-22. Claim 23 uses the generic term “stem cell” as it relates to differentiating induced pluripotent stem cells into enteric neural crest cells for administration in subjects with enteric neuropathy. Claim 27 uses the generic term “stem cell” as it relates to differentiating induced pluripotent stem cells into enteric neural crest cells for administration in subjects with enteric neuropathy. The same generic scope for “stem cell” is present in each of the dependent claims, i.e. 32-38. The specification provides antecedent basis for “stem cell” as it relates to induced pluripotent stem cells being differentiated into enteric neural crest cells [pg. 2, line 31]. Applicant’s specification states “the ENCCs are differentiated from pluripotent stem cells in vitro. In some embodiments, the stem cells are embryonic stem (ES) cells or induced pluripotent stem cells (iPSCs) [Summary of the invention ¶ 1]. The specification further gives examples where the stem cells are from the cell line LiPSC-GR1.1, i.e. an induced pluripotent stem cell. Throughout Applicant’s specification, the stem cells referred to are induced pluripotent stem cells. However, “stem cells” covers generically all stem cells, including mesenchymal stem cells. It is known that mesenchymal stem cells are multipotent and are only capable of differentiating into a limited range of cells such as osteoblasts, chondrocytes, or adipocytes. Although these cells can be induced to exhibit neuronal like features, mesenchymal stem cells do not naturally possess the developmental potential to generate ectodermal derivatives such as neural crest cells. They lack the ability to differentiate into enteric neurons or glia. Because of this, a person of ordinary skill would not understand how to differentiate mesenchymal stem cells into enteric neural crest cells given the art teaches the use of pluripotent stem cells. Given this, the generic term “stem cells” as it relates to stem cells being differentiated into enteric neural crest cells and the absence of teaching of how to accomplish this with stem cells other than pluripotent type stem cells, an Artisan would not understand that Applicant is in possession of the full scope of “stem cells” as it relates to differentiating stem cells into enteric neural crest cells based on the Applicant’s specification referencing only pluripotent type stem cells. Claim Rejections - 35 USC § 112 07-30-02 AIA 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. 07-34-01 Claims 4, ,10, 14, 22, 26, 32, and 35-36 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. 07-34-10 AIA Regarding claim 4 , the phrase "such as" renders the claim indefinite because it is unclear whether the limitations following the phrase are part of the claimed invention. See MPEP § 2173.05(d). 07-34-05 AIA Claim 10 recites the limitation " cells " in Line 1 . There is insufficient antecedent basis for this limitation in the claim. Claim 15 recites “administering one or more additional agents selected from the group consisting of…”. It is unclear if the additional agent is being administered to the subject or combined with the cell composition prior to the composition being administered. Claim 22 recites “immune system surveillance”. As written, it is unclear by what the Applicant means. Is this referencing adaptive immunity, innate immunity, or antigen presenting pathways such as HLA class I and II, or is it referring to inflammatory signaling or cytokine responses, or a combination of some or all. A person of ordinary skill would not understand what is meant by this term. Claim 26 recites “mixture of cells are positive for HNK-1, positive for expression p75, negative for expression Oct4 and expression of Nanog”. However, the claim also states “wherein the expression of HNK-1, p75, Oct4 and/or Nanog…”. These statements are inconsistent making the claim unclear as to what is being claimed. Is Oct4 and Nanog to be present or not? A person of ordinary skill would not understand whether Oct4 and Nanog are required to be present or not. 07-34-05 AIA Claim 32 recites the limitation " medium " in Line 2 . There is insufficient antecedent basis for this limitation in the claim. Claim Rejections - 35 USC § 102 07-06 AIA 15-10-15 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. 07-07-aia AIA 07-07 The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action: A person shall be entitled to a patent unless – 07-08-aia AIA (a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention. 07-15 AIA Claim s 1, 2, 5, 6, 18, 19, 20, 21, and 23-25 are rejected under 35 U.S.C. §102( a)(1 ) as being anticipated by Chang et al. [Induced pluripotent stem cell-derived enteric neural crest cells repopulate human aganglionic tissue-engineered intestine to from key components of the enteric nervous system, J Tissue Eng ., 2020] . Regarding claim 1, Chang et al. teaches a method to treat an enteric neuropathy [Abstract] comprising administering enteric neural crest cells (ENCCs) to a muscular wall of the intestine in a subject in need thereof [Figure 1 & Figure 2], wherein the ENCCs are differentiated from stem cells [Abstract]. Regarding claim 2, Chang et al. teaches the method of claim 1, wherein the enteric neuropathy is Hirschsprung’s Disease (HD) [Introduction ¶ 1]. Regarding claim 5, Chang et al. teaches the method of claim 1, wherein the entire intestine is aganglionic [Abstract]. Regarding claim 6, Chang et al. teaches the method of claim 1, wherein the cells are administered by injection [Figure 1]. Regarding claim 18, Chang et al. teaches the method of claim 1, wherein the stem cells are induced pluripotent stem cells or embryonic stem cells [HIO and ENCC generation]. Regarding claim 19, Chang et al. teaches the method of claim 18, wherein the induced pluripotent stem cells comprise the cell line of LiPSC-GR1.1 [HIO and ENCC generation ¶ 1]. Regarding claim 20, Chang et al. teaches the method of claim 1, wherein the cells are allogeneic to said subject [HIO and ENCC generation ¶ 1]. Regarding claim 21, Chang et al. teaches the method of claim 1, wherein the cells are genetically modified [HIO and ENCC generation ¶ 1]. Regarding claim 23, Chang et al. teaches enteric neural crest cells differentiated from stem cells in vitro and a cryopreservative [HIO and ENCC generation ¶ 1. Furthermore, LiPSC-GR1.1 is shipped frozen where it is typically cryopreserved in a protective solution such as DMSO]. Regarding claim 24, Chang et al. teaches wherein the stem cells comprise induced pluripotent stem cells or embryonic stem cells [HIO and ENCC generation ¶ 1]. Regarding claim 25, Chang et al. teaches the composition of claim 24, wherein the induced pluripotent stem cells comprise the cell line LiPSC-GR1.1 [HIO and ENCC generation ¶ 1] . 07-15 AIA Claim 26 is rejected under 35 U.S.C. §102( a)(1 ) as being anticipated by Huang et al. [Characterization and transplantation of enteric neural crest cells from human induced pluripotent stem cells, Nature: Molecular Psychiatry , 2016] . Regarding claim 26, Huang et al. teaches a neurosphere [hiPSC-NCSCs can colonize aganglionic human gut tissue in vitro ¶ 1] comprising a heterogenous mixture of cells, wherein the mixture of cells are positive for expression of HNK-1, positive for expression p75, negative for expression of Oct4 and negative for expression Nanog, wherein the expression of HNK-1, p75, Oct 4 and/or Nanog is from one cell or each marker is expressed individually or in combination on different cells [Derivation of enteric-like neurons from hiPSC-NCSCs in vitro ¶ 1] . Claim Rejections - 35 USC § 103 07-06 AIA 15-10-15 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. 07-20-aia AIA 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. 07-23-aia AIA 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. 07-21-aia AIA Claim s 3-4, 7-17, and 27-38 are rejected under 35 U.S.C. §103 as being unpatentable over Chang et al. [Induced pluripotent stem cell-derived enteric neural crest cells repopulate human aganglionic tissue-engineered intestine to from key components of the enteric nervous system, J Tissue Eng ., 2020], in view of Moore [Hirschsprung Disease: current perspectives, Taylor & Francis Group , 2015], in view of Meneghel et al. [Cryopreservation as a key element in the successful delivery of cell-based therapies – a review, Front. Med ., 2020], in view of McKeown et al. [Exposure to GDNF enhances the ability of enteric neural progenitors to generate an enteric nervous system, Stem Cell Reports , 2017], in view of Fattahi et al. [Deriving human ENS lineages for cell therapy and drug discovery in Hirschsprung disease, Nature , 2016], in view of Menendez et al. [Wnt signaling and a SMAD pathway blockage direct the differentiation of human pluripotent stem cells to multipotent neural crest cells, Cell Biology , 2011], in view of Allison [Immunosuppressive therapy in transplantation , Nursing Clinics of North America , 2016], in view of Lee et al. [Divergent effects of Wnt/Beta-catenin signaling modifiers on the preservation of human limbal epithelial progenitors according to culture condition, Nature: Scientific Reports , 2017] . Chang et al. teaches each and every element for claims 1 and 2. However, Chang et al. does not specifically cite total intestinal agangliosis form of HD or a long segment form of HD as stated in claim 3. However, Moore, discussing the known characteristics of Hirschsprung disease, discloses long segment agangliosis being associated with HD, as well as ultra-long segment aganglionosis, also known as Zuelzer’s disease [Familial transmission ¶ 2]. This disclosure coupled with the disclosure in Chang et al. where the abstract mentions the absence of intestinal nerves, it would have prima facie obvious to a person of ordinary skill prior to the filing of the claimed invention to modify the systems and methods of Chang et al. that discloses using enteric neural crest cells derived from pluripotent stem cells for treating enteric neuropathy, e.g. HD, where the treatment focused on total agangliosis or long-segment agangliosis. Given this, there is a reasonable expectation of success that an artisan would recognize the teachings of both Chang et al. and Moore where enteric neural crest cells derived from pluripotent cells, natural or induced, could be differentiated into enteric neural crest cells for the treatment of total or long-segment agangliosis resulting from HD. For claim 4 where the method of claim 1 is used to treat enteric neuropathy chosen from a Markush listing, Moore teaches that Hirschsprung disease, although varies considerably, is known to classically affect the rectum and sigmoid where the aganglionosis extends proximally for short and long segments depending on phenotypic expression [Pathology ¶ 2]. For claim 7 where the cells are administered to two or more locations, Although Chang et al. discloses the method of administering enteric neural crest cells by injection, Chang et al. does not specifically disclose method of administering enteric neural crest cells to two or more locations of the intestine. However, administering stem cells or enteric neural crest cells to two or more locations along the intestine would have been a matter of routine optimization using known methods, given a person of ordinary skill in the art would recognize that the intestine is an elongated organ and that multiple-site injection administration would be necessary in order to improve cell distribution, engraftment, and homing of the enteric neural crest cells. Additionally, for claims 8, 9, and 10 where the administration of the cells involve bilateral injections every 1 to 3 centimeters, a person of ordinary skill would also understand that in order to maximize efficacy and homing of the enteric neural crest cells, given the elongated nature of the intestines, that multiple injections would be required for each section of the intestine in order for the neural crest cell base to cover the intestinal wall based where the number of injections would be determined in the amount of agangliosis present in the subject. This would depend further whether the area affected was due to total agangliosis, short-form agangliosis, long-form agangliosis, or even ultra long-form agangliosis, a person of ordinary skill would recognize that in order for the administration of enteric neural crest cells needed to cover the entire affected area of the intestine would require both multiple injections along the length of the intestine at predetermined lengths, as wells as bilateral injections for treating the entire circumference of the intestine. Additionally, targeting the muscular wall of the intestine would also be obvious to a person of ordinary skill given that restoring neurons in the myenteric plexus region would be critical for restoring peristalsis and/or coordinated muscle contraction. Additionally, Chang et al. discloses multiple injection sites shown as scaffold seeding in part (a) in Figure 1. For claim 11 and claim 12 where the method of claim 8 is made up of a dose comprising a particular cellular concentration, Chang et al. discloses the concentration of 1 x 10 5 cells [Surgical procedures ¶ 1]. However, cell concentration selection also represents a matter of routine optimization using known methods. A person of ordinary skill in the art would know to adjust cell density or dose to achieve a therapeutic benefit based on considerations such as target tissue, delivery method, and treatment efficacy. For claim 13, Chang et al. discloses pluripotent stem cell-derived enteric neural crest cells were introduced into an engineered small intestine implant as a cell suspension. Additionally, the cell suspension included an acceptable carrier where the cells were prepared in a liquid injection medium [Figure 1]. For claim 14 where the cells are cryopreserved, Meneghel et al., discloses that cryopreservation is a key enabling technology for use in regenerative medicine allowing for secure extended cell storage of prepared cell preparations [Abstract]. For claim 15 where additional agents, such as pH buffering agent, is administered, McKeown et al., discussing GDNF and its role in generating an enteric nervous system, discloses that GDNF resulted in a 14-fold increase in nuerosphere volume and a 12-fold increase in cell number following co-culture with embryonic gut or transplantation into the colon of postnatal mice. Additionally, cells derived from GDNF-treated neurospheres showed a 2-fold increase in the distance migrated compared with controls [Abstract]. Given this, there is a reasonable expectation of success that a person of ordinary skill in the art would recognize the benefits of utilizing GDNF as a migration agent given its ability increase migration in enteric neural progenitor cells. Therefore, it is prima facie obvious to a person of ordinary skill in the art prior to the filing of the claimed invention to modify the systems and methods of Chang et al. with the additional teachings of McKeown et al. where a person of ordinary skill would recognize the benefits of utilizing GDNF as a migration agent in order to better facilitate homing of the enteric neural crest cells in allowing these cells to migrate and develop into an enteric nervous system as it relates to Hirschsprung disease. For claim for claim 16 where the migration promotion agent includes GDNF, again McKeown discloses the use of GDNF as a plausible migration agent given its success in increases both volume and migration distance with enteric neuronal progenitor cells [Abstract]. For claim 17 where the subject has been clinically immunosuppressed prior to administration, Allison, discussing immunosuppressive therapy in transplantation, discloses that immunosuppressive medications are given in order to modulate the recipients immune response to the donor organ, tissue, and/or cells [Key Points]. This involves both induction, e.g. prior to surgery, and maintenance therapy [Key Points]. For claim 27 where a method of producing enteric neural crest cells contains a number of steps, Fattahi et al., discussing methods for culturing and deriving human enteric nervous system cell lineages, including neural crest cells, discloses expanding the cells in suspension [In vitro differentiation of ENC to enteric neurons ¶ 1] contacting the stem cells with FGF2 and retinoic acid [Neural crest induction ¶ 1, Main ¶ 1], and further isolating the resultant cell lines [Abstract]. However, Fattahi et al. is silent on the use of an inhibitor of SMAD and a WNT pathway activator. For the SMAD and WNT signaling, Menendez et al., discussing ways to differentiate human pluripotent stem cells into neural crest cells, discloses that concurrent SMAD inhibition combined with activation of WNT signaling markedly increased p75 cells, a known cellular marker for neural crest cells [Results: Activation of the Wnt Pathway Redirects Neural Progenitors toward a Neural Crest Fate ¶ 1]. For claim 28 where the stem cells are induced pluripotent stem cells, Menendez et al. discloses the use of induced pluripotent stem cells for deriving multipotent neural crest cells [Introduction ¶ 3]. For claim 29 wherein the iPSCs comprise the cell line LiPSC-GR1.1, Chang et al. discloses the use of LiPSC-GR1.1 as the cell line being used to derive neural crest cells [HIO and ENCC generation ¶ 1]. Given this, there is a reasonable expectation of success that a person of ordinary skill in the art would combine the teachings of Fattahi et al. with the teachings of both Menendez et al. and Chang et al. where an artisan, for the purpose of deriving neural crest cells, would chose a known method using established cell lines, such as LiPSC-GR1.1, where the cell line was suspended in culture where the pluripotent stem cells were exposed to FGF2 and retinoic acid for a period of time to allow differentiation. There is also a reasonable expectation of success that a person of ordinary skill in the art would also use the additional teachings of Menendez et al. by recognizing the importance of inhibiting SMAD while simultaneously activating the WNT signaling pathway to further direct differentiation towards a neural crest cell. Given this, it would have been prima facie obvious to a person of ordinary skill in the art prior to the filing of the claimed invention to modify the systems and method of Faffahi et al. with the further teachings of both Menendez et al. and Chang et al. For claim 30 where the inhibitor can be SB431542, Menendez et al. discloses the use of SM431542 as an inhibitor capable of blocking SMAD [Results: Activation of the Wnt Pathway redirects neural progenitors towards a neural crest fate ¶ 3]. For claim 31 where the WNT activator is CHIR99021, Lee et al., discussing WNT signaling modifiers, discloses that CHIR99021 is a known as a primary inducer of the WNT signaling canonical pathway [Abstract]. For claim 32, the same analysis for claim 27 applies. For claim 33 where the where the isolated enteric neural crest cells are cryopreserved prior to administration, the same analysis for claim 14 applies. For claim 34, the same analysis for claim 27 applies. For claim 35 where the enteric neural crest cell culture is substantially homogenous, Huang et al. discloses using FACS to isolate p75 and HNK-1 dual positive cells where these cells represented approximately 90% among the differentiated HDF-hiPSCs [Generation of NCSCs from hiPSCs ¶ 2]. For claim 36 where the enteric neural crest cells comprise a heterogenous cell population, Huang et al., similar to the above statement, discloses a heterologous cell population that contains dual positive p75 and HNK-1 cells among other cell types [Id.]. For claim 37 where a pharmaceutical composition containing enteric neural crest cells, the same analysis for claim 13 is applicable. For claim 38 containing a method for treating an enteric neuropathy by injecting the enteric neural crest cells to a muscular wall of the intestine to a subject in need thereof, the same analysis for claim 1 coupled with the analysis for claim 34 is applicable. The Supreme court has acknowledged: When a work is available in one field of endeavor, design incentives and other market forces can prompt variations of it, either in the same field or a different one. If a person of ordinary skill can implement a predictable varition..103 likely bars its patentability …if a technique has been used to improve one device, and a person of ordinary skill in the art would recognize that it would improve similar devices in the same way, using the technique is obvious unless its actual application is beyond that person’s skill. A court must ask whether the improvement is more than the predictable use of prior-art elements according to their established functions… … the combination of familiar elements according to known methods is likely to be obvious when it does no more than yield predictable results (see KSR International Co. v. Teleflex Inc ., 82 USPQ2d 1385 U.S. 2007) emphasis added. In KSR Int'l Co. v. Teleflex Inc. , 550 U.S. 398 (2007), the Supreme Court reaffirmed "the conclusion that when a patent 'simply arranges old elements with each performing the same function it had been known to perform' and yields no more than one would expect from such an arrangement, the combination is obvious." Id. at 417 (quoting Sakraida v. Ag Pro, Inc ., 425 U.S. 273,282 (1976)). The Supreme Court also emphasized a flexible approach to the obviousness question, stating that the analysis under 35 U.S.C. § 103 "need not seek out precise teachings directed to the specific subject matter of the challenged claim, for a court can take account of the inferences and creative steps that a person of ordinary skill in the art would employ." Id. at 418; see also id. at 421 ("A person of ordinary skill is... a person of ordinary creativity, not an automaton."). From the teachings of the references, it is apparent that one of ordinary skill in the art would have had a reasonable expectation of success in producing the claimed invention. 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, as evidenced by the references, especially in the absence of evidence to the contrary. Conclusion No claims allowed. Any inquiry concerning this communication or earlier communications from the examiner should be directed to JOHN DAVID MOORE whose telephone number is (703)756-1887. The examiner can normally be reached M-F 8-5. 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, Tracy Vivlemore can be reached on 571-272-2914. 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. /JOHN DAVID MOORE/Examiner, Art Unit 1638 /Tracy Vivlemore/Supervisory Primary Examiner, Art Unit 1638 Application/Control Number: 18/559,119 Page 2 Art Unit: 1638 Application/Control Number: 18/559,119 Page 3 Art Unit: 1638 Application/Control Number: 18/559,119 Page 4 Art Unit: 1638 Application/Control Number: 18/559,119 Page 5 Art Unit: 1638 Application/Control Number: 18/559,119 Page 6 Art Unit: 1638 Application/Control Number: 18/559,119 Page 7 Art Unit: 1638 Application/Control Number: 18/559,119 Page 8 Art Unit: 1638 Application/Control Number: 18/559,119 Page 9 Art Unit: 1638 Application/Control Number: 18/559,119 Page 10 Art Unit: 1638 Application/Control Number: 18/559,119 Page 11 Art Unit: 1638 Application/Control Number: 18/559,119 Page 12 Art Unit: 1638 Application/Control Number: 18/559,119 Page 13 Art Unit: 1638 Application/Control Number: 18/559,119 Page 14 Art Unit: 1638 Application/Control Number: 18/559,119 Page 15 Art Unit: 1638 Application/Control Number: 18/559,119 Page 16 Art Unit: 1638 Application/Control Number: 18/559,119 Page 17 Art Unit: 1638 Application/Control Number: 18/559,119 Page 18 Art Unit: 1638 Application/Control Number: 18/559,119 Page 19 Art Unit: 1638 Application/Control Number: 18/559,119 Page 20 Art Unit: 1638 Application/Control Number: 18/559,119 Page 21 Art Unit: 1638