DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 11/20/2025 has been entered.
Claim Status
Claim 1 is amended.
Claims 14-21 are withdrawn.
Claims 1-13 are under examination.
Response to Amendment
The edited ground of rejection below addresses the deficiencies raised by Applicant with respect to the amended claims.
Edited rejections necessitated by claim amendment.
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.
Claims 1-3 are rejected under 35 U.S.C. 103 as being unpatentable over Stebbins et al ( Science Advances, 2019, cited on IDS), in view of Miller et al (Biology open,2017), as evidenced by Schroeter et al (Nature, 1998) and Aihara et al (International journal of developmental biology) .
Stebbins et al teach a method of generating the neural crest stem cells (NCSCs), the embryonic precursor to forebrain pericytes, from Human pluripotent stem cells (hPSCs), and then differentiating NCSCs to brain pericyte–like cells.(See the abstract).
Regarding claims1-3, Stebbins et al teach a method for differentiating p75-NGFR+HNK-1+ neural crest stem cells into a population of mural cell lineage. Stebbins method produces cells that express NG2+PDGFRβ+⍺SMAlow , indicating brain pericyte-like cells. Stebbins et al do not mention that the cells also express RGS5. In the instant application, the whole genome transcriptome of Stebbins cells was compared to the primary in vivo brain pericyte, indicating that the cells produced by Stebbins method also express RGS5, albeit at a lower level than their in vivo counterpart. (See Fig.6A, instant application). Stebbins et al teach that culturing the NCSCs in E6 medium supplemented with 10% fetal bovine serum (FBS) for 9 days is sufficient to direct the NCSCs pacification toward brain pericyte-like cells. (See Fig.2C-E).
Stebbins et al teach the requirement for serum to induce the differentiation of NCSCs into mural cell linage, but do not teach the requirement of Notch signaling or culturing the cells in serum-free medium.
Miller et al teach the use of in ovo electroporation system to insert a tetracycline-inducible NotchΔE construct (encoding a constitutively active mutant of mouse Notch1) into the genome of chicken cranial neural crest cell precursors, and the activation of NotchΔE expression by doxycycline injection at embryonic day 4. Miller et al teach the constitutive activation of Notch1 is sufficient in vivo to drive the neural crest-derived frontonasal mesenchyme to a perivascular/mural cell fate. This reads on claim 1 step (a). (See the abstract). It should be noted that, in the material and methods section “ Electroporation constructs ” page 323, Miller et al refers to the protocol of Sato et al as source reference for constructing the NotchΔE construct, who subsequently refers to the protocol of Schroeter et al as source reference for constructing the NotchΔE construct (See material and methods section” Expression vectors”, page 899). According to Schroeter et al, the NotchΔE construct encodes for the intracellular (NICD) and the transmembrane domains of the mouse Notch1, but lacking the entire extracellular domain. ( See Schroeter et al Fig.2A). Hence, Miller et al use the intracellular domain of the mouse Notch 1 (Uniport accession # Q01705). This reads on claim 2 step (i). In addition, Miller et al also state that while “the mural cells originate from either mesoderm or cranial neural crest, regardless of their origin, Notch signaling is necessary for their formation”. (See abstract). Furthermore, Miller et al demonstrate that constitutively activating Notch1 was sufficient in vivo to drive both somite cells, and neural crest-derived frontonasal mesenchyme, to a perivascular (i.e. mural cells) cell fate. Clearly, the teachings of Miller encompass motivation, suggestion, and teachings, as well as experimental basis for one skilled in the art to readily envision activating Notch signaling in the cells of Stebbins to produce mural cells, and with a high expectation of success.
In other words, it would have been prima facie obvious to one with ordinary skill in the art at the time the invention was filed to combine Stebbins and Miller’s teachings to increase Notch expression in NCSCs in order to induce their differentiation into a mural cell lineage. Stebbins et al teach differentiating the NCSCs into a brain-like pericyte by culturing the cells in E6 medium supplied with 10% FBS, but fail to teach increasing the expression of Notch. Miller et al teach the use of N1ICD is sufficient to drive the neural crest-derived frontonasal mesenchyme to mural cell fate. Thus, one would have been motivated to combine the teachings of Stebbins and Miller to use N1ICD to drive the differentiation of neural crest cells into mural cell fate, because Notch signaling is known to play critical roles in stem cells by regulating cell fate determination and differentiation, and Notch1 signaling is required for neural crest-derived mural cell formation as taught by Miller. Combining prior art elements according to known methods to yield predictable results. See MPEP 2143 (I)(A).
The teachings of Stebbins et al and Miller et al are set forth above.
It is noted that the in vivo system of Miller also contains serum and does not teach the requirement for any media composition. Therefore, an ordinary skill in the art, upon reviewing the teachings of Miller et al, can easily extrapolate that the sustained activation of the Notch signaling is sufficient enough to drive the differentiation of the neural crest cells into a mural fate without the need for serum or specific medium. Stebbins et al, on the other hand, demonstrate that NCSCs may be maintained in a chemically defined serum-free medium, wherein the media is a serum-free medium supplemented with GSK3 B inhibitor, a TGFB inhibitor, a BMP type I receptor inhibitor and FGF2 ( known as E6-CSFD). Stebbins et al, specifically teach how to use E6-CSFD medium to expand NCSCs prior their differentiation to different lineages. ( See sections “MACS of NCSCs” and “ NCSC lineage differentiations” on page 11). Stebbins et al also report that NCSCs expanded in E6-CSFD medium retain the neural crest marker. ( See right column on page 6- line 44-46).
On the other hand, Aihara et al state that “ serum contains variable and undefined amounts of soluble growth and differentiation factors, as well as extracellular components such as fibroblast growth factors (FGFs), bone morphogenetic proteins (BMPs), fibronectin, and laminin; thus the presence of serum may cloud our understanding of the mechanisms of cell differentiation”. (See introduction,2nd column, page 1287). As a result, Aihara et al developed a protocol using defined serum-free medium to differentiate neural crest cells from mouse embryonic stem cells (mES) in the presence of FGF-2 and BMP-4. (See Fig.3 G-H) . Aihara et al also teach that culturing neural crest cells in a serum-free medium supplemented with different combinations of factors can shift their differentiation into different cell lineages. For example, Aihara et al teach culturing the neural crest in ESF5 medium with FGF-2 and BMP-4 drives differentiation into glial cell lineage. On the other hand, culturing neural crest cells in ESF5 medium with FGF-2, BMP-4, and PDGF causes them to differentiate into Schwann cells. (See Fig.5 A-C). However, when neural crest cells were cultured in DMEM medium supplemented with 10% fetal calf serum, smooth muscle actin positive cells appeared (See.Fig.5E). Taken together, an ordinary skill in the art ,upon reviewing the teachings of Aihara, would turn away from using a serum supplemented medium during cell differentiation because serum contains variables and undefined number of factors, resulting in variable results and reducing reproducibility, and would instead turn to a chemically defined serum-free medium to ensure reproducibility in research.
Collectively, Stebbins et al, demonstrate an advantage for culturing NCSCs in E6-CSFD medium (i.e. a chemically defined serum-free medium) so that cells reach the appropriate cell density before differentiation. It should be noted that E6-CSFD medium reads on the medium recited in step (b) of instant claim 1. Aihara et al teach culturing NCSCs in serum-free medium and clearly suggest using a defined serum-free culture system during cell differentiation to clarify the mechanism of induction, as well as to ensure the reproducibility of the results.( See left column, 1st paragraph, on page 1293). Miller et al demonstrate that constitutively activating Notch1 in vivo is sufficient to drive both somite cells, and neural crest-derived frontonasal mesenchyme, to a perivascular (i.e. mural cells) cell fate.
In other words, claim 1 is combining prior art elements according to known methods to yield predictable results, namely the predictable result is the production of mural cells from NCSCs by constitutively activating Notch signaling. Because an ordinary skill in the art would find it instructive that Miller et al teach that constitutively activating Notch1 is sufficient to drive the differentiation of neural crest-derived frontonasal mesenchyme into a perivascular (i.e. mural cells) cell fate. Also, a person of ordinary skill in the art, who had reviewed Miller and Aihara, could have come across Stebbins and immediately noticed the strong possibility of using the E6-CSFD medium of Stebbins would have the predictable result of propagating NCSCs prior to their differentiation, and one would then use N1ICD to drive the differentiation of neural crest cells into mural cell fate, as taught by Miller, because Notch1 signaling is required for neural crest-derived mural cell formation. One would also be motivated to drive the differentiation of NCSCs into mural fate by activating Notch signaling while culturing the cells in the defined serum-free medium of Stebbins (i.e. E6-CSFD) because one would know that the removal of serum would generate more reproducible results. Combining prior art elements according to known methods to yield predictable results. See MPEP 2143 (I)(A).
Claims 4,5,7-13 are rejected under 35 U.S.C. 103 as being unpatentable over Stebbins et al ( Science Advances, 2019, cited on IDS), in view of Miller et al (Biology open,2017) and Aihara et al (International journal of developmental biology) as applied to claims 1-3 above, and further in view of Rusanescu et al ( US 2016/0032244 A1) and Chen et al (Stem cells and development, 2012).
The teachings of Stebbins, Miller, and Aihara are set forth above. Stebbins in view of Miller and Aihara render obvious claims 1-3. Stebbins, Miller, and Aihara do not teach the use of human Notch3 intracellular domain (N3ICD) or the human Notch1 intracellular domain (N1ICD).
Regarding claims 4 and 13, following the discussion above, Stebbins and Miller do not teach the human Notch3 intracellular domain (N3ICD). Rusanescu et al teach the use of a viral construct that encodes for the human N3ICD in mouse Neuro-2a cells induces these cells to differentiate into neurons. Rusanescu et al also teach culturing these cells in serum free medium. Although the N3ICD transfected in this example was of human origin, the expressed N3ICD was capable of performing its intended action of inducing the differentiation of Neuro-2a cells of mouse origin. (See example 4, paragraphs [ 0110, and 0111], and SEQ ID # 2). This reads on SEQ ID 2 of instant claim. Rusanescu et al also suggest the use of stem cells transfected with a vector that expresses N3ICD may be introduced in a mammal for the purpose of differentiating into various cell types within that mammal. Therefore, it would have been prima facie obvious to one with ordinary skill in the art at the time the invention was filed to combine the teachings of Stebbins, Miller, and Rusanescu to use the human N3CID to induce NCSCs into a population of mural cell lineage. Stebbins et al teach differentiating the NCSCs into a brain-like pericyte by culturing the cells in E6 medium supplied with 10%FBS. Miller et al teach Notch signaling is required for neural crest-derived mural cell formation. Rusanescu et al teach inducing N3ICD expression into Neuro-2a cells cultured in serum-free medium induces them to differentiate into neurons. Thus, one would have been motivated to combine the teachings of Stebbins, Miller, and Rusanescu to use the human N3ICD to drive the differentiation of neural crest cells into mural cell fate, because Notch signaling is known to play critical roles in stem cells by regulating cell fate determination and differentiation, Notch3 is also prominently expressed in brain mural cells, and Notch signaling is required for neural crest-derived mural cell formation. Combining prior art elements according to known methods to yield predictable results. See MPEP 2143 (I)(A).
Regarding claim 5, following the discussion above, Stebbins, and Miller do not teach the human Notch1 intracellular domain (N1ICD). Chen et al teach the use of a construct that encodes for the human N1ICD to drive the differentiation of the cardiosphere derived progenitor cells into a smooth muscle cell lineage. ( See abstract, Fig.3, and the Materials and Methods section “ Plasmid construct and generation of recombinant adenoviruses”).
Therefore, it would have been prima facie obvious to one with ordinary skill in the art at the time the invention was filed to combine the teachings of Stebbins, Miller, and Chen to use the human N1CID to induce NCSCs into a population of mural cell lineage. Stebbins et al teach differentiating the NCSCs into a brain-like pericyte by culturing the cells in E6 medium supplied with 10%FBS. Miller et al teach the use of mouse N1ICD is sufficient to drive the neural crest-derived frontonasal mesenchyme to mural cell fate. Chen et al teach the use of human N1ICD is sufficient to drive the cardiosphere derived progenitors to smooth muscle cells lineage. Thus, one would have been motivated to combine the teachings of Stebbins, Miller, and Chen to use the human N1ICD to drive the differentiation of the human neural crest cells into mural cell fate, because Notch signaling is known to play critical roles in stem cells by regulating cell fate determination and differentiation, and Notch1 signaling is required for neural crest-derived mural cell formation as taught by Miller. Combining prior art elements according to known methods to yield predictable results. See MPEP 2143 (I)(A).
Regarding claims 7 and 12, Stebbins et al teach that culturing the NCSCs in E6 medium supplemented with 10% fetal bovine serum (FBS) for 9 days is sufficient to direct the NCSCs specification toward brain pericyte-like cells. (See Fig.2C-E). This reads on claim 7. Stebbins et al also teach that following the 9 days culturing of NCSCs in E6 and 10%FBS generated at least 90% brain pericyte-like cells. This reads on claim 9. ( See Fig1J).
Regarding claims 8 and 9, In the instant application, the whole genome transcriptome of Stebbins cells was compared to the primary in vivo brain pericyte, demonstrating that the cells produced by Stebbins method also express RGS5, but at a lower level than their in vivo counterpart (See Fig.6A, instant application). This reads on claim 8. The method of Stebbins also produces cells that express KCNJ8 and ABCC9. (See Fig.2B). This reads on claim 9.
Regarding claims 10 and 11, Stebbins et al teach culturing hPSC in E6-CSFD medium for 15 days to produce p75-NGFR+HNK-1+NCSC. This reads on claim 10. (See Fig. 1A-C and J, and Fig. S1, B- E). Stebbins et al also teach the magnetic sorting for the p75-NGFR+ and the replating of p75-NGFR+ to produce an enriched population of p75-NGFR+NCSC. This reads on claim 11. ( See Fig. 1F,G and J).
Claim 6 is rejected under 35 U.S.C. 103 as being unpatentable over Stebbins et al ( Science Advances, 2019, cited on IDS), in view of Miller et al (Biology open,2017), as evidenced by Schroeter et al (Nature, 1998), further in view of Aihara et al (International journal of developmental biology) and Taqvi et al (Journal of Biomedical Materials Research Part A, 2006).
The teachings of Stebbins et al ,Miller et al , and Aihara are set forth above. Stebbins, Miller, and Aihara do not teach the use of beads coated with Notch ligands.
Taqvi et al teach the use of a microbead coated with Notch ligand to drive stem cells differentiation into the T-cell lineage. ( See the abstract).
Regarding claim 6, Taqvi et al teach co-culturing of bone marrow-derived hematopoietic stem cells (BMHSCs) with stromal cells in the presence of magnetic microbeads coated with notch ligand DLL4 is sufficient to drive the BMHSCs into T-lymphocyte cell lineage. (See Fig.4).
Therefore, it would have been prima facie obvious to one with ordinary skill in the art at the time the invention was filed to combine the teachings of Stebbins, Miller, Aihara, and Taqvi to use Notch activated NCSCs cultured in serum-free medium in the presence of beads coated with Notch ligand to induce them to differentiate into a population of mural cell lineage. Stebbins et al teach differentiating the NCSCs into a brain-like pericyte by culturing the cells in E6 medium supplied with 10%FBS. Miller et al teach the use of mouse N1ICD is sufficient to drive the neural crest-derived frontonasal mesenchyme to mural cell fate. According to Aihara, serum contains variable and undefined amounts of soluble growth and differentiation factors, thus the presence of serum may cloud our understanding of the mechanisms of cell differentiation. Taqvi et al teach the use of culturing the BMHSCs in the presence of microbead coated with Notch ligand drive them to differentiate into the T-cell lineage. Thus, one would have been motivated to combine the teachings of Stebbins, Miller, Aihara, and Taqvi to activate Notch signaling in NCSCs by culturing them in serum free medium containing beads coated with Notch ligand to drive their differentiation into mural cell fate, because Notch signaling is known to play critical roles in stem cells by regulating cell fate determination and differentiation, and Notch signaling is required for neural crest-derived mural cell formation as taught by Miller. Combining prior art elements according to known methods to yield predictable results. See MPEP 2143 (I)(A). Taken together, the invention as claimed is unpatentable over the work of prior art.
Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
Claims 1-3,9-11 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-4 of U.S. Patent No. 11,643,636 B2, hereinafter 636’, in view of Stebbins et al ( Science Advances, 2019), Miller et al (Biology open,2017), as evidenced by Schroeter et al (Nature, 1998) and Aihara et al (International journal of developmental biology).
Regarding claims 1-3,9-11, the claims of ’636 teach a method of generating neural crest stem cells (NCSCs) from human pluripotent stem cells (hPSCs), and then differentiating NCSCs to brain pericyte–like cells. Specifically, the method first involves differentiating hPSCS into NCSCs by culturing the cells in E6-CSFD medium for 15 days. The cells are then sorted for NCSCs and replated in E6-CSFD medium to produce enriched population of NCSCs. After that, the propagated NCSCs are cultured in E6 medium supplemented with 10% fetal bovine serum (FBS) for 9 days to direct their specification toward brain pericyte-like cells.
It is noted that the claims of 636’ teach the requirement for serum to induce the differentiation of NCSCs into mural cell linage but do not teach the requirement for Notch signaling or culturing the cells in serum-free medium.
The teachings of Stebbins, Miller and Aihara are set forth above.
Therefore, it would have been prima facie obvious to one with ordinary skill in the art to combine the teachings of 636’, Stebbins, Miller, and Aihara to increase the expression of Notch signaling in NCSCs to induce their differentiation into mural lineage. Because an ordinary skill in the art would find it instructive that Miller et al teach that constitutively activating Notch1 is sufficient to drive the differentiation of neural crest-derived frontonasal mesenchyme into a perivascular (i.e. mural cells) cell fate. Also, a person of ordinary skill in the art, who had reviewed Miller and Aihara, could have come across Stebbins and 636’ and immediately noticed the strong possibility of using the E6-CSFD medium, of Stebbins and 636’ would have the predictable result of propagating NCSCs prior to their differentiation, and one would then use N1ICD to drive the differentiation of neural crest cells into mural cell fate, as taught by Miller, because Notch1 signaling is required for neural crest-derived mural cell formation. One would also be motivated to drive the differentiation of NCSCs by activating Notch signaling while culturing the cells in the defined serum-free medium of Stebbins and 636’ (i.e. E6-CSFD) because one would know that the removal of serum would generate more reproducible results.
Response to Arguments
Applicant's arguments filed 11/20/2025 have been fully considered but they are not
persuasive.
Applicants argue that Stebbins et al teach the requirement of serum ( rather than a serum-free medium as required by claim 1) in the culture for the differentiation of mural cell lines. Applicants also argue that the method of Miller inherently occurs in serum-containing medium rather than a serum-free medium, and doesn’t teach any media composition as required by claim 1.
Examiner's Response to Traversal: Applicant's arguments have been carefully
considered but are not found persuasive. This is because Applicants attack Stebbins et al and Miller et al individually, notwithstanding the fact that claim 1 is rejected over Stebbins et al in view of Miller, and Aihara. Applicants are reminded that arguments against a single reference do not obviate a 103 based on multiple references and so these arguments that Stebbins et al do not teach the differentiation of neural crest stem cells into mural cells under a serum-free conditions are not persuasive. One cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981 ); In re Merck & Co., Inc., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986). This is because "[T]he test for obviousness is what the combined teachings of the references would have suggested to [a PHOSITA]." In re Mouttet, 686 F.3d 1322, 1333, 103 USPQ2d 1219, 1226 (Fed.Cir. 2012).
Applicants also argue that Aihara teaches that the differentiation of neural crest cells into other cell populations depends on the culture media composition, and does not teach which culture media should be used to differentiate neural crest cells into mural cells. Hence, Applicants argue that the combination of references do not disclose or suggest each
element of the claims
Examiner's Response to Traversal: Applicant's arguments have been carefully
considered but are not found persuasive. This is because obviousness may be established by combining or modifying the teachings of the prior art to produce the claimed invention where there is some teaching, suggestion, or motivation to do so found either in the references themselves or in the knowledge generally available to one of ordinary skill in the art. See In re Fine, 837 F.2d 1071, 5USPQ2d 1596 (Fed. Cir. 1988), In re Jones, 958 F.2d 347, 21 USPQ2d 1941 (Fed. Cir. 1992), andKSR International Co. v. Teleflex, Inc., 550 U.S. 398, 82 USPQ2d 1385 (2007). In this case, and as discussed above, a person of ordinary skill in the art, who had reviewed Miller and Aihara, could have come across Stebbins and immediately noticed the strong possibility of using the E6-CSFD medium of Stebbins would have the predictable result of propagating NCSCs prior to their differentiation, and one would then use N1ICD to drive the differentiation of neural crest cells into mural cell fate, as taught by Miller, because Notch1 signaling is required for neural crest-derived mural cell formation. One would also be motivated to drive the differentiation of NCSCs into mural fate by activating Notch signaling while culturing the cells in the defined serum-free medium of Stebbins (i.e. E6-CSFD) because one would know that the removal of serum would generate more reproducible results.
Conclusion
No claim is allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to FATIMAH KHALAF MATALKAH whose telephone number is (703)756-5652. The examiner can normally be reached Monday-Friday,7:30 am-4:30 pm 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, 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.
/FATIMAH KHALAF MATALKAH/Examiner, Art Unit 1638
/Tracy Vivlemore/Supervisory Primary Examiner, Art Unit 1638