Prosecution Insights
Last updated: April 19, 2026
Application No. 17/429,540

BIOFUNCTIONALIZED HYDROGEL FOR CELL CULTURE

Final Rejection §103§112§DP
Filed
Aug 09, 2021
Examiner
ZHU, JIANJIAN
Art Unit
1631
Tech Center
1600 — Biotechnology & Organic Chemistry
Assignee
VANDERBILT UNIVERSITY
OA Round
4 (Final)
62%
Grant Probability
Moderate
5-6
OA Rounds
3y 8m
To Grant
99%
With Interview

Examiner Intelligence

Grants 62% of resolved cases
62%
Career Allow Rate
46 granted / 74 resolved
+2.2% vs TC avg
Strong +82% interview lift
Without
With
+82.0%
Interview Lift
resolved cases with interview
Typical timeline
3y 8m
Avg Prosecution
70 currently pending
Career history
144
Total Applications
across all art units

Statute-Specific Performance

§101
3.3%
-36.7% vs TC avg
§103
32.9%
-7.1% vs TC avg
§102
18.0%
-22.0% vs TC avg
§112
27.2%
-12.8% vs TC avg
Black line = Tech Center average estimate • Based on career data from 74 resolved cases

Office Action

§103 §112 §DP
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 . DETAILED ACTION Amendments In the reply filed 12/22/2025, Applicant has amended claims 30 and 57, and newly canceled claims 47 and 58-59. Claim Status Claims 30, 36, 46, 48 and 51-57 are pending and are considered on the merits. Information Disclosure Statement The information disclosure statements (IDS) submitted on 10/09/2025 and 03/12/2026 are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statements are being considered by the examiner. The corresponding signed and initialed PTO forms 1449 have been mailed with this action. Withdrawn Claim Rejections - 35 USC § 112(b) The prior rejection of claims 30, 36, 46, 48 and 51-57 under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite by reference to a relative term is withdrawn in light of amendment to claim 30. Withdrawn Claim Rejections - 35 USC § 103 The prior rejection of claims 30, 36, 46, 48 and 51-57 under 35 U.S.C. 103 set forth in the prior Office action mailed on 08/22/2025 is withdrawn in light of Applicant’s amendment to recite new limitations such as a step of embedding the plurality of cells (including a neuron) in a biomaterial, which is not discussed in the prior rejection. New Claim Rejections - 35 USC § 103 The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action: A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, 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. Claims 30, 36, 46, 48, 51-54 and 57 are rejected under 35 U.S.C. 103 as being unpatentable over Lancaster et al., (Nature. 2013;501(7467):373-379) in view of Oksdath et al., (APL Bioeng. 2018;2(4):041501, p. 1-19), Zhou et al., (ACS Appl. Mater. Interfaces. 2018, 10, 8993-9001. Prior art of record) and Lim et al., (ACS Biomater. Sci. Eng. 2017, 3, 776-781, S1-S8. Prior art of record). With respect to claim 30, Lancaster develops a human pluripotent stem cell-derived three-dimensional organoid culture system, termed cerebral organoids (e.g., abstract), thus teaches the preamble a method of culturing a plurality of cells. In regard to culturing iPSCs to provide a plurality of cells including a neuron, Lancaster teaches human iPS cell line SC101A-1 cells (“Method” p. 1, left col., para 2) and reprogrammed patient iPS cells (“Method” p. 2, left col.) are cultured in neural induction media to form neuroepithelial tissues (“Method” p. 1, section “Cerebral organoid culture conditions”, see Fig 1A, Day 0 – Day 11). Lancaster teaches the method leads to rapid development of brain tissue, requiring only 8-10 days for the appearance of neural identity (p. 373, right col, para 2. It is noted that this brain tissue likely includes a neuron). Thus, Lancaster suggests culturing iPSCs to provide a plurality of cells including a neuron. In regard to embedding the plurality of cells in a biomaterial of a crosslinked hydrogel, Lancaster teaches on day 11 of the protocol, neuroepithelial tissues were transferred to droplets of Matrigel (i.e., a hydrogel) and these droplets were allowed to gel at 37 °C (i.e., to crosslink) (“Method” p. 1, section “Cerebral organoid culture conditions”), thus teaches embedding the plurality of cells in a biomaterial of a crosslinked hydrogel. In regard to culturing the cells embedded in the biomaterial for at least 3 weeks, Lancaster teaches the tissue droplets are cultured in stationary condition for 4 days (see Fig 1A, Day 11 – Day 15) and then transferred to a spinning bioreactor (“Method” p. 1, section “Cerebral organoid culture conditions”) and cultured for defined brain regions to form at 30 days (p. 373, right col, para 2) and for more developed tissues at 75 days (p. 376, left col, para 3). Thus, Lancaster teaches culturing the cells embedded in the biomaterial for at least 3 weeks. However, Lancaster is silent on the biomaterial consisting of a crosslinked hydrogel comprising gelatin, and a peptide comprising SEQ ID NO: 1 chemically attached to the hydrogel. Nevertheless, Lancaster teaches the protocol focuses on improving growth conditions and providing the environment necessary for intrinsic cues to influence development, and teaches the biomaterial Matrigel is to provide a scaffold for more complex tissue growth (p. 373, right col, para 2). Oksdath summarizes biomaterial-based strategies that optimize stem cell-derived brain organoid growth (e.g., title, abstract and Fig 3). Oksdath acknowledges that current protocols for brain organoid growth use commercially available Matrigel, which poorly reflects the composition of the brain tissue (at any stage) and serves mostly as a scaffold/mechanical support (see, for example, Lancaster et al., 2013. It is noted that this is the prior art Lancaster recited above). Matrigel also presents other limitations including a limited capacity for the user to vary their mechanical properties and biochemical composition to match those of the ECM in the normal brain tissue, and these unmatched properties have been shown to be critical for neuronal differentiation, growth, and development of neural connections in 3D cultures (p. 6, last para – p. 7, para 1). Oksdath suggests that the synthetic ECM provides new ways to finely control the physical, chemical, and biological characteristics of the organoid microenvironment, with advantages: (i) the controlled attachment of biomolecules in the gel structure, (ii) the possibility of modulating the mechanical properties of the materials, and (iii) the degree of degradability of the material (Fig. 3) (p. 7, para 3 “Synthetic materials to control the 3D “homogenous” organoid environment”, also see p. 13, last 2 para). Zhou teaches a synthetic ECM mimic GelMA for neuron differentiation and culturing (p. 8895, left col, para 2.1, p. 8997, last para, and Fig 7), which is crosslinked by a UV laser (p. 8995, left col, para 2.2). Zhou teaches gelatin methacrylate (GelMA) possesses excellent biocompatible and biodegradable properties because it contains many RGD and matrix metalloproteinase sequences (e.g., abstract) and furthermore, modifying substrate properties with functional molecules (such as DA) can induce the differentiation of NSCs into neurons for diverse neuroregenerative applications (p. 8995, left col, para 2). Thus, Zhou teaches a synthetic crosslinked hydrogel comprising gelatin (GelMA) and suggests modifying the GelMA with functional molecules. Lim teaches an N-Cadherin-derived peptide Ac-HAVDIGGGC (that is 100% identical to the instant SEQ ID NO: 1) that is covalently tethered into a hydrogel (PEGDMA) for neuron differentiation and culturing (abstract, p. S2, “Materials” and p. 777, left col, last para, see Fig 1A), thus teaches a peptide chemically attached to a hydrogel wherein the peptide comprises SEQ ID NO: 1. Lim teaches the NCAD-derived peptide is a superior candidate for inclusion into a biomaterial as it is easier to evenly disperse through matrices due to its smaller size and it can stimulate similar cellular receptors and pathways compared to that by native NCAD which plays an important role in the development and homeostasis of the central nervous system (p. 776, last para and p. 776, left col, last para). Therefore, it would have been obvious for one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of culturing iPSC-derived plurality of cells including a neuron by embedding the cells in a crosslinked Matrigel disclosed by Lancaster, by substituting the Matrigel with a synthetic ECM mimic GelMA hydrogel as suggested by Oksdath and taught by Zhou, and further combining a peptide comprising SEQ ID NO: 1 chemically attached to the hydrogel as suggested by Zhou and taught by Lim with a reasonable expectation of success. Since Oksdath acknowledges Matrigel used by Lancaster has limitations such as not able to vary biochemical composition to match normal brain tissue and instead suggests using synthetic ECM having the controlled attachment of biomolecules (see above), since Zhou teaches a synthetic crosslinked hydrogel GelMA possesses excellent biocompatible and biodegradable properties and can be modified with functional molecules (see above), and since Lim teaches the NCAD-derived peptide (having SEQ ID NO: 1) is a superior candidate for inclusion into a biomaterial as it is easier to evenly disperse through matrices and it can stimulate similar cellular receptors and pathways of native NCAD which plays an important role in the development and homeostasis of the central nervous system (see above), one of ordinary skill in the art would have had a reason to substitute the Matrigel with a synthetic GelMA and to further combine an NCAD-derived peptide chemically attached to GelMA in order to take advantages of the biocompatible and biodegradable properties of GelMA and the controlled attachment of biomolecules (the NCAD-derived peptide) to stimulate native NCAD pathway for neuronal differentiation and culture. PNG media_image1.png 369 351 media_image1.png Greyscale Furthermore, the reasonable expectation of success is supported by the fact that both Zhou and Lim use a hydrogel having a methacrylate (MA) group and Lim teaches “a cysteine attached to a three glycine spacer was added to the C-terminus of the N-terminus acetylated HAVDI peptide to allow covalent tethering into the hydrogel” (p. 777, left col, last para, see modified Fig 1 of Lim and modified Fig 1 of Zhou, note that the HAVDI peptide is chemically attached to the methacrylate group in Lim), thus one ordinary skill in the art would have had a reasonable expectation of success in chemically attaching the HAVDI peptide of Lim to the GelMA hydrogel of Zhou. In regard to the limitation “wherein active synapses between the plurality of cells are increased by at least 80% after about 21 days of culturing compared to active synapses between the plurality of cells at the onset of culturing”, it must be noted that this wherein clause does not recite an active step in the claimed method, but only the results of the culturing the plurality of cells embedded in the biomaterial for at least 3 weeks as suggested by Lancaster in view of Oksdath, Zhou and Lim. MPEP 2111.04 (I) states a whereby clause (or a wherein clause) “in a method claim is not given weight when it simply expresses the intended result of a process step positively recited.” Therefore, this wherein clause does not provide any patentable weight in determining patentability of the claimed method. Nevertheless, Lancaster teaches culturing the plurality of cells embedded in the biomaterial for at least 3 weeks results in neuronal activity, evidenced by calcium oscillations (p. 376, left col, para 5, see Fig 4e for calcium oscillation and see Fig 1C-Fig 4 for neuron marker TUJ1). Zhou teaches that enhanced TUJ1 staining was notable on GelMA scaffold over time (p. 8997, last section 3.4 “Neural Differentiation of NSCs on 3D Scaffolds”, see Fig 7F for neuron differentiation and connection in GelMA). Lim teaches the neuronal markers TUJ1 (Figure 4B) and microtubule associated protein 2 (MAP2) (Figure 4C) exhibited maximal expression in hNSC cultured on hydrogels containing 577 μM HAVDI (p. 778, right col, para 3, see Fig 4, compared to hydrogels with lower concentrations of the peptide). Thus, Lancaster, Zhou and Lim suggest that culturing the plurality of cells including neurons in the biomaterial would have likely resulted in increased active synapses formation by the recited percentage. With respect to claim 36, Lancaster teaches a cerebral organoid culture system (see e.g., Fig 1), thus teaches the plurality of cells is a cerebral organoid. With respect to claims 46 and 48, as stated supra, Lim teaches “a cysteine attached to a three glycine spacer was added to the C-terminus of the N-terminus acetylated HAVDI peptide to allow covalent tethering into the hydrogel” (p. 777, left col, last para, see modified Fig 1 above), thus teaches the peptide being attached to the hydrogel at the C-terminal end in claim 46. Lim teaches the HAVDI peptide is derived from N-cadherin protein and can stimulate similar cellular receptors (i.e., comprises an extracellular epitope) compared to native NCAD (e.g., p. 777, left col, para 1), thus teaches the peptide comprises an extracellular epitope of a cadherin protein in claim 48. Accordingly, one of ordinary skill in the art would have chosen the peptide sequence taught by Lim in the method of Lancaster in view of Oksdath, Zhou and Lim since Lim has reduced to practice the sequence of the HAVDI peptide with proven function in enhancing neurite growth. With respect to claims 51-54 directed to the hydrogel being crosslinked by a crosslinker, Zhou teaches the GelMA hydrogel is crosslinked by a crosslinker acrylic acid group which undergoes a cross-linking reaction after UV exposure in the presence of an I2959 photoinitiator (e.g. see Fig 1 for an acrylic anhydride crosslinker and Fig 2 for the cross-linking reaction), thus Zhou teaches the hydrogel being crosslinked by a crosslinker in claim 51, the hydrogel being crosslinked by a UV-light activated crosslinker in claim 52, the crosslinker comprising a -C(CH3)=CH2 group in claim 54 (see Fig 2B for the structure of the crosslinker acrylic acid base). It is noted that claim 53 is directed to optional limitations that do not carry patentable weight, and thus is rejected in the same way as of claim 51. With respect to claim 57, as stated supra, Lancaster teaches the plurality of cells is a cerebral organoid (see e.g., abstract and Fig 1). Lancaster teaches the brain organoid has laminar pattering of cortical layers (see e.g., Fig 1C, Fig 2, Fig 3, Fig 4 and Fig 6). Accordingly, one of ordinary skill in the art would have immediately expected that the method of Lancaster in view of Oksdath, Zhou and Lim would have resulted in the brain organoid to have laminar patterning of cortical layers. Hence, the claimed invention as a whole was prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention in the absence of evidence to the contrary. Response to Traversal: Applicant' s arguments filed on 12/22/2025 are acknowledged. Applicant first argues that the present application surprisingly found that biomaterials as claimed were able to provide physical and biochemical cues and replace the synaptogenic role of astrocytes and yield more viable cells (specification [0079] and [0082]), and a pronounced increase in the expression of postsynaptic terminal markers on neurons, improved synaptic connectivity and widespread neural network formation (Example 4 and Figs 9-10), and facilitate maturation of iPSC-derived neurons on a functional level (Remarks, p. 5-6). Applicant’s arguments have been fully considered but they are not persuasive. As a first matter, as stated supra, Lancaster teaches a biomaterial (Matrigel) enables neuronal differentiation and maturation in a brain organoid (e.g., abstract, see Fig 4e for calcium oscillation as a functional readout and see Fig 1C-Fig 4 for expression of neuron marker TUJ1, and see Fig 1C, Fig 2, Fig 3, Fig 4 and Fig 6 for laminar pattering of cortical layers). Zhou teaches that enhanced TUJ1 staining was notable on GelMA scaffold over time (p. 8997, last section 3.4 “Neural Differentiation of NSCs on 3D Scaffolds”, see Fig 7F for neuron differentiation and connection in GelMA). Lim teaches the neuronal markers TUJ1 (Figure 4B) and microtubule associated protein 2 (MAP2) (Figure 4C) exhibited maximal expression in hNSC cultured on hydrogels containing 577 μM HAVDI (p. 778, right col, para 3, see Fig 4, compared to hydrogels with lower concentrations of the peptide). Thus, Lancaster, Zhou and Lim suggest that culturing the plurality of cells including neurons in the biomaterial would have likely resulted in increased active synapses formation by the recited percentage. Accordingly, the biomaterials as claimed function as expected based on the teachings of Lancaster, Zhou and Lim. Furthermore, MPEP 716.02(d), states that unexpected results must be commensurate in scope with the claimed invention. In the instant case, the purported “surprising” results presented by the Applicant were studies performed by (1) differentiating human iPSCs into cortical glutamatergic neurons and culturing for 70-100 days, (2) embedding the neurons into a GelMA-Cad hydrogel, and (3) culturing the neurons within the GelMA-Cad hydrogel for 21 days, which is not commensurate in scope with the claimed method of culturing iPSCs to provide a plurality of cells including a neuron, embedding the plurality of cells in a biomaterial consisting of a crosslinked hydrogel comprising gelatin (i.e., any hydrogel comprising gelatin and any other components) with the peptide chemically attached, and culturing the plurality of cells in the biomaterial for at least 3 weeks. Applicant further argues that Zhou and Lim teach culturing neural stem cells and iPSC-derived neural stem cells but nowhere does these references teach or suggest culturing iPSC-derived neural cells with the method as claimed. The culturing methods of Zhou and Lim are silent on any synaptic activity of its neural stem cells, thus suggest their cultured cells were unable to form mature neural networks. In fact, Zhou and Lim do not characterize their neural stem cells besides neurite length and gene expression. As such, one of skill in the art would fail to arrive at a method of culturing cells as claimed and would be especially unable to predict the surprising culturing benefits provided by the claimed method (Remarks, p. 6). Applicant’s arguments have been fully considered and they are persuasive. Specifically, Zhou and Lim are silent on the amended limitations of embedding the plurality of cells within the biomaterial and culturing for at least 3 weeks. Therefore, the prior rejection has been withdrawn. However, as necessitated by amendment, a new ground of rejection has been made over Lancaster in view of Oksdath, Zhou and Lim as discussed above. Specifically, Lancaster teaches a method of embedding the plurality of cells including a neuron within a biomaterial (Matrigel) and culturing the cells for at least 3 weeks. Furthermore, Lancaster characterizes the cells being mature neurons by (1) expression of marker gene TUJ1, (2) structure of the brain organoid having laminar pattering of cortical layers, and (3) at functional levels having calcium oscillations. Claim 55 is rejected under 35 U.S.C. 103 as being unpatentable over Lancaster et al., (Nature. 2013;501(7467):373-379) in view of Oksdath et al., (APL Bioeng. 2018;2(4):041501, p. 1-19), Zhou et al., (ACS Appl. Mater. Interfaces. 2018, 10, 8993-9001. Prior art of record) and Lim et al., (ACS Biomater. Sci. Eng. 2017, 3, 776-781, S1-S8. Prior art of record), as applied to claim 30 above, and further in view of Lee et al., (Materials 2016, 9, 797, p. 1-13. Prior art of record). With respect to claim 55 directed to the gelatin comprising porcine skin gelatin, Zhou teaches the gelatin (type A, Sigma-Aldrich) is used (p. 8995, left col, para 2.1). However, Lancaster, Oksdath, Zhou and Lim do not specifically teach this type A gelatin of Zhou from Sigma-Aldrich is from porcine skin. Lee teaches a method for making bioink using GelMA (abstract). Lee teaches GelMA can be classified into two types: type A GelMA (a product from acid treatment) and type B GelMA (a product from alkali treatment) (abstract). Lee teaches to synthesizing type A GelMA, gelatin type A (Bloom 175 g, porcine skin, Sigma-Aldrich, St. Louis, MO, USA) is used (p. 2, para 2.1), thus teaches the type A gelatin from Sigma-Aldrich is from porcine skin. Therefore, it would have been obvious for one of ordinary skill in the art before the effective filing date of the claimed invention to have appreciated the type A gelatin from Sigma-Aldrich used in the method of Lancaster in view of Oksdath, Zhou and Lim would have been from porcine skin as taught by Lee. One of ordinary skill in the art would have had a reason to use porcine skin gelatin because Zhou has reduced to practice using a commercially available gelatin derived from porcine skin that contains many RGD and matrix metalloproteinase sequences and possesses excellent biocompatible and biodegradable properties even after it is photocured under UV light. Hence, the claimed invention as a whole was prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention in the absence of evidence to the contrary. Response to Traversal: Applicant' s arguments filed on 12/22/2025 are acknowledged and have been discussed above. Claim 56 is rejected under 35 U.S.C. 103 as being unpatentable over Lancaster et al., (Nature. 2013;501(7467):373-379) in view of Oksdath et al., (APL Bioeng. 2018;2(4):041501, p. 1-19), Zhou et al., (ACS Appl. Mater. Interfaces. 2018, 10, 8993-9001. Prior art of record) and Lim et al., (ACS Biomater. Sci. Eng. 2017, 3, 776-781, S1-S8. Prior art of record), as applied to claim 30 above, and further in view of Wu et al., (Bioscience Reports. 2019 January; 39: BSR20181748, p. 1-9. Prior art of record). Claim 56 is directed to the biomaterial having a stiffness of about 800 Pa to about 5 kPa. However, Lancaster, Oksdath, Zhou and Lim are silent on the stiffness of the GelMA biomaterial. Nevertheless, Okadath summarizes the stiffness of the mice and human brain regions in which human gray matter has a stiffness of 3.1 kPa and white matter 2.7 kPa (see e.g., Fig 2). Wu teaches the stiffness of GelMA influences the outgrowth of a representative neuron cell line (abstract). Wu teaches the 5% GelMA hydrogel has the Young’s modulus of 3.08 KPa (within the claimed range of about 800 Pa to about 5 kPa), and the representative neuronal cells on the 5% GelMA hydrogel show the highest adhesion rate (p. 4 Results section 1 and section 3, see Fig 4 for Young’s muduli and Fig 6A for cell adhesion on GelMA hydrogels). Wu teaches the stiffness of GelMA can be modified by adjusting the concentration of GelMA and this helps the design and optimization of tissue engineering scaffolds for nerve regeneration (abstract). Therefore, it would have been obvious for one of ordinary skill in the art before the effective filing date of the claimed invention to have modified the method of Lancaster in view of Oksdath, Zhou and Lim, by substituting with 5% GelMA hydrogel having a stiffness of about 800 Pa to about 5 kPa as suggested by Okadath and taught by Wu with a reasonable expectation of success. Since Okadath summarizes the stiffness of human gray matter being 3.1 kPa and white matter 2.7 kPa (see e.g., Fig 2), and since Wu teaches 5% GelMA hydrogel with a stiffness of 3.08 KPa results in the highest cell adhesion rate (p. 4 Results section 3, see Fig 6A), one of ordinary skill in the art would have had a reason to use 5% GelMA hydrogel as taught by Wu with the claimed stiffness in order to mimic the human brain tissue environment and to enhance the neuron adhesion to the hydrogel to improve the culturing method. Hence, the claimed invention as a whole was prima facie obvious to a person of ordinary skill before the effective filing date of the claimed invention in the absence of evidence to the contrary. Response to Traversal: Applicant' s arguments filed on 12/22/2025 are acknowledged and have been discussed above. Withdrawn Provisional Double Patenting Rejection The prior rejection of claims 30, 36, 46, 48 and 51-57 on the ground of nonstatutory double patenting as being unpatentable over claims 1-15 of copending Application No. 18/268,009 in view of Daviaud et al., (eNeuro. 2018, 5(6) e0219-18.2018, p. 1-18. Prior art of record) is withdrawn in light of Applicant’s amendment to claim 30 to recite new limitations of embedding the plurality of cells in the biomaterial and culturing the plurality of cells embedded in the biomaterial for at least 3 weeks, which are not made obvious over the cited application claims in view of Daviaud. Response to Traversal: Applicant' s arguments filed on 12/22/2025 are acknowledged. Applicant requests to hold the rejection in abeyance until there is allowable subject matter in the present application (Remarks, p. 7). Applicant’s arguments have been fully considered but they are not persuasive. However, as stated supra, the prior rejection has been withdrawn in light of Applicant’s amendment to claim 30 to recite new limitations. Pertinent Reference Prior art that is considered relevant to Applicant’s invention but not relied upon for rejection: Fan et al., (ACS Appl. Mater. Interfaces. 2018, 10, 17742-17755) disclose a method of culturing iPSC-derived neural stem cells (iNSCs) comprising culturing iPSCs to provide iNSCs, embedding the iNSCs in a 3D gelatin methacrylate (GelMA) hydrogel, and culturing the iNSCs embedded in the GelMA both in vitro (for 7 days) and in vivo (for 6 weeks) such that the active synapses are increased (marked by synaptophysin, see Figure S3, the same marker used by the instant invention). Conclusion Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a). A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any extension fee pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the date of this final action. No claims are allowed. Examiner Contact Information Any inquiry concerning this communication or earlier communications from the examiner should be directed to Jianjian Zhu whose telephone number is (571)272-0956. The examiner can normally be reached M - F 8:30AM - 4PM (EST). 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, James Douglas (Doug) Schultz can be reached on (571) 272-0763. 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. /JIANJIAN ZHU/Examiner, Art Unit 1631 /JAMES D SCHULTZ/Supervisory Patent Examiner, Art Unit 1631
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Prosecution Timeline

Aug 09, 2021
Application Filed
Sep 04, 2024
Non-Final Rejection — §103, §112, §DP
Dec 04, 2024
Response Filed
Feb 26, 2025
Final Rejection — §103, §112, §DP
Jun 04, 2025
Request for Continued Examination
Jun 06, 2025
Response after Non-Final Action
Aug 12, 2025
Non-Final Rejection — §103, §112, §DP
Dec 22, 2025
Response Filed
Apr 06, 2026
Final Rejection — §103, §112, §DP (current)

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Prosecution Projections

5-6
Expected OA Rounds
62%
Grant Probability
99%
With Interview (+82.0%)
3y 8m
Median Time to Grant
High
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