Notice of Pre-AIA or AIA Status
The present application is being examined under the pre-AIA first to invent provisions.
DETAILED OFFICE ACTION
This Office Action is in response to the papers filed on 10 April 2025.
CLAIMS UNDER EXAMINATION
Claims 184-189, 191-193, 195-197, 199-200 and 206-209 are pending and have been examined on their merits.
PRIORITY
The Applicant claims priority to Provisional Application 61/126,802, filed on 06 May 2008, and Provisional Application 61/190,282, filed on 26 August 2008.
The ‘802 Provisional Application does not provide support for basic fibroblast growth factor or the claimed concentration of said growth factor. A search of the document indicates the terms “fibroblast growth factor”, “basic fibroblast growth factor”, “FGF-2” (also known as basic fibroblast growth factor) or “bFGF” do not appear in the text. The document does not provide support for all of the limitations recited in claims 193 and 197. The document does not provide support for all of the growth factors recited in claims 189 and 191 (i.e. bFGF).
Claim 207 recites 25-100 ng/ml BMP-4 in step a. The ‘802 Application provides support for 50 ng/ml BMP-4 (line 3 and line 29 of page 39). It does not provide support for the range recited in claim 207 or a concentration of 25 ng/ml as recited in claim 208.
Claim 207 recites 25-100 ng/ml VEGF in step a. The ‘802 Application provides support for 50 ng/ml VEGF165 (line 3 and line 27 of page 39). It does not provide support for the entire range recited in claim 207. Provisional Application ‘282 provides support for growing human ES cells in the presence of 25-100 ng/ml VEGF 165 ([0136]). The earliest priority for claims 207-208 is 26 August 2008. Provisional Application ‘282 Application does not provide support for the limitations recited in claims 193.
WITHDRAWN REJECTIONS
Claims 184 and 185 have been amended to require 25-100 ng/ml VEGF. The rejection of claims 184, 186, 188-189, 191-192 and 195-197 under pre-AIA 35 U.S.C. 103(a) as being unpatentable over Pick et al. in view of Rajesh et al. and Crawford et al. is withdrawn because the references due not teach this limitation.
NEW GROUNDS OF REJECTION
Claims 184 and 185 have been amended to require 25-100 ng/ml VEGF. New grounds of rejection have been necessitated by claim amendment.
Claim Rejections - 35 USC § 103
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claims 184-186, 188, 191-195, 197, 199-200, 206-207 and 209 are rejected under pre-AIA 35 U.S.C. 103(a) as being unpatentable over Lu et al. (previously cited; Generation of functional hemangioblasts from human embryonic stem cells. Nat Methods. Epub 07 May 2007) in view of Rajesh et al. (previously cited; In Vitro Differentiation Of Hematopoietic Cells From Primate Embryonic Stem Cells. US2008/0108044 08 May 2008 with benefit Provisional application 60/857756 08 November 2006).
Lu et al. generate progenitor cells with both hematopoietic and vascular potential from hES cells using a two-step procedure with different supplements under fully serum-free conditions (first paragraph of page 2).
Human embryonic (hES) cells are cultured on ultra-low dishes in serum-free medium with the addition of BMP-4 and VEGF165 (50 ng/ml each) to induce hemangioblast precursor (mesoderm) formation for 48 hours (see page 7, last paragraph). Because Lu teaches (a) of claim 184, it would inherently induce the formation of embryoid bodies.
48 hours later, Lu removes half the medium and added fresh medium with the same final concentrations of BMP-4 and VEGF (e.g., 50 ng/ml each), plus SCF, Tpo and FLT3 ligand (20 ng/ml each; R&D Systems) to expand the hemangioblast and its precursor.
Lu collects EBs and dissociated them and prepared single-cell suspension (last paragraph of page 7). To expand hemangioblasts, the single-cell suspension derived from differentiation of 2–5 × 105 hES cells is mixed with 2 ml of BGM containing 1.0% methylcellulose in Isocve’s MDM, 1–2% bovine serum albumin, 0.1 mM 2-mercap-toethanol, 10 μg/ml rh-insulin, 200 μg/ml iron saturated human transferrin, 20 ng/ml rh-GM-CSF, 20 ng/ml rh-IL-3, 20 ng/ml rh-IL-6, 20 ng/ml rh-G-CSF, 3–6 units/ml rh-Epo, 50 ng/ml rh-SCF, 50 ng/ml rh-VEGF and 50 ng/ml rh-BMP-4, with or without 50 ng/ml of Tpo and FL. Therefore Lu cultures in a second serum free medium that contains TPO and FL.
The deficiency of Lu is that it does not teach 20-40 ng/ml bFGF in the second medium.
Rajesh teaches a method of generating hematopoietic lineage cells and hemangioblasts from primate (human and non-human primates) embryoid bodies (EBs). The method is summarized as including the steps of forming embryoid bodies from embryonic stem cells, and then culturing the EBs under serum-free conditions in a differentiation medium that is supplemented periodically with fibroblast growth factor (FGF) in an amount sufficient to yield differentiated hematopoietic lineage cells and hemangioblasts ([0012]). Rajesh teaches the FGF is FGF-2 (hence, basic fibroblast growth factor) in some embodiments ([0014]). Rajesh teaches a concentration from above 0.5 ng/ml to about 50 ng/ml for human cells ([0019]).
It would have been obvious to use bFGF in the second medium taught by Lu. One would so since Lu teaches a method of producing hemangioblasts using embryoid bodies and Rajesh teaches adding bFGF to generate hemangioblasts from embryoid bodies. It would have been obvious to use 20-40 ng/ml bFGF since Lu uses human cells and Rajesh teaches using 0.5 ng/ml to about 50 ng/ml for human cells. One would have had a reasonable expectation of success since Rajesh teaches bFGF can be used to generate hemangioblasts. One would have expected similar results since both references are directed to methods of differentiating human embryonic stem cells. Therefore claim 184 is rendered obvious.
Regarding independent claim 185: In addition to the limitations recited in claim 184, the claim recites disaggregating embryoid bodies.
The teachings of Lu are reiterated. Lu collects EBs and dissociated them and prepared single-cell suspension (last paragraph of page 7). The deficiency of Lu is that it does not teach 20-40 ng/ml bFGF.
The teachings of Rajesh are reiterated.
It would have been obvious to use bFGF in the second medium taught by Lu. One would so since Lu teaches a method of producing hemangioblasts using embryoid bodies and Rajesh teaches adding bFGF to generate hemangioblasts from embryoid bodies. It would have been obvious to use 20-40 ng/ml bFGF since Lu uses human cells and Rajesh teaches using 0.5 ng/ml to about 50 ng/ml for human cells. One would have had a reasonable expectation of success since Rajesh teaches bFGF can be used to generate hemangioblasts. One would have expected similar results since both references are directed to methods of differentiating human embryonic stem cells. Therefore claim 185 is rendered obvious.
Lu teaches human embryonic stem cells (supra). Therefore claim 186 is rejected. Lu cultures in VEGF and BMP-4 for 48 hours (supra). Therefore claim 188 is included in this rejection. Lu teaches the second medium comprises EPO, SCF, BMP and VEGF (supra). Therefore claim 191 is included in this rejection. Lu teaches EPO in the second medium (supra). Therefore claim 192 is included in this rejection. Lu teaches culturing in a medium comprising methylcellulose in the second step as recited above (supra). Therefore claims 195-196 are rejected. Lu teaches “fully serum-free conditions” (see first paragraph of page 2). Therefore claim 197 is rejected. Lu teaches hematopoietic and endothelial lineage development (last paragraph of page 2). Therefore claims 199-200 are included in this rejection.
Lu teaches cells are plated in the second medium and cultured for 4-5 days (see first paragraph of page 8). Blast colonies are produced (same cited section). Lu teaches blast colonies can be used for CFU assays, endothelial cell development or in vivo transplantation (see page 8, last two paragraphs). Lu is silent regarding the number of cells obtained after 4-5 days of culture. It would have been obvious to culture the cells taught by Lu to obtain at least 1x106 hemangio-colony forming cells. One would do so to produce enough cells for the in vitro and in vivo studies taught by Lu. Therefore claim 206 is rendered obvious as claimed.
Lu teaches 50 ng/ml BMP-4 (supra). Therefore claim 207 is rejected.
Lu teaches VEGF 165 (supra). Therefore claim 209 is rejected.
Therefore Applicant’s Invention is rendered obvious as claimed.
Claim 193 is rejected under pre-AIA 35 U.S.C. 103(a) as being unpatentable over Lu in view of Rajesh as applied to claim 184 above, and further in view of Watanabe et al. (previously cited; A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nature Biotechnology. published online 27 May 2007. Volume 25 (6) pages 682-686).
Claim 184 is rejected on the grounds set forth above. The teachings of Lu and Rajesh are reiterated. Lu teaches a method of differentiating human embryonic stem cells. The art teaches forming EBs. EBs are dissociated.
Lu is silent regarding the use of a ROCK inhibitor in either medium.
Watanabe teaches poor survival of human embryonic stem (hES) cells after cell dissociation is an obstacle to research, hindering manipulations (Abstract). hES cells are vulnerable to apoptosis upon cellular detachment and dissociation. They undergo massive cell death particularly after complete dissociation. Thus, hES cells are difficult, if not impossible, to use in dissociation culture, which is important for such procedures as differentiation induction (page 682, left column, first paragraph). Watanabe teaches Y-27632 has a protective ability which enables human embryonic stem cells to survive and differentiate following dissociation (page 682, right column, first full paragraph). The art teaches the inhibitor has this effect even in serum free conditions (see Abstract).
It would have been obvious to use a ROCK inhibitor in the serum free medium used by Lu. One would have been motivated to do so since Lu uses dissociated and differentiated human embryonic stem cells that have been and Watanabe teaches ROCK inhibitor protects apoptosis of said cells when they are dissociated. One would use the same inhibitor to prevent cell death. One would have had a reasonable expectation of success using a ROCK inhibitor with pluripotent stem cells since, as set forth above, they are interpreted to be equivalent the human embryonic stem cells. One would have expected similar results since both references use the same cell type in serum free medium. Therefore claim 193 is included in this rejection.
Therefore Applicant’s Invention is rendered obvious as claimed.
Claim 187 is rejected under pre-AIA 35 U.S.C. 103(a) as being unpatentable over Lu in view of Rajesh as applied to claim 184 above, and further in view of Takahashi et al. (previously cited; Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors. Cell 131, 861–872, November 30, 2007) as evidenced by UCLA (definition: Induced Pluripotent Stem Cells (iPS).
Claim 184 is rejected on the grounds set forth above. The teachings of Lu and Rajesh are reiterated. While Lu discloses the use of human embryonic stem cells, Lu is silent regarding the use of “human induced pluripotent stem cells”.
As defined by UCLA, induced pluripotent stem cells are “are derived from skin or blood cells that have been reprogrammed back into an embryonic-like pluripotent state”.
Takahashi et al. demonstrate the generation of iPS cells from adult human dermal fibroblasts (see Summary). The art teaches “human iPS cells were similar to human embryonic stem (ES) cells in morphology, proliferation, surface antigens, gene expression, epigenetic status of pluripotent cell-specific genes, and telomerase activity. Furthermore, these cells could differentiate into cell types of the three germ layers in vitro and in teratomas” (see Summary). Said cells are “comparable to human ES cells in their differentiation potential in vitro and in teratomas” (see page 861, right column, second full paragraph). Takahashi teaches the use of human embryos faces ethical controversies that hinder the applications of human ES cells. In addition, it is difficult to generate ES cells. One way to circumvent these issues is to induce pluripotent status in somatic cells by direct reprogramming (page 861, Introduction left column bridging first paragraph of right column).
It would have been obvious to the person of ordinary skill in the art at the time the invention was made to substitute a human induced pluripotent stem cell for the human embryonic stem cell taught by Lu. A human induced pluripotent stem cell is a stem cell that is in an “embryonic-like pluripotent state”. Takahashi teaches induced pluripotent stem cells are similar to human embryonic stem cells in morphology, proliferation, surface antigens, gene expression, epigenetic status of pluripotent cell-specific genes, and telomerase activity. Therefore the cited cells appear to be functional equivalents. KSR B teaches it is rational to substitute one known, equivalent element for another to obtain predictable results. The skilled artisan would use a cell that has been induced to behave the same was as human embryonic stem cells. Takahashi teaches the use of human embryos faces ethical controversies that hinder the applications of human ES cells. In addition, it is difficult to generate ES cells. One way to circumvent these issues is to induce pluripotent status in somatic cells by direct reprogramming. One would be motivated to use reprogrammed cells which are easier to generate and which are not hindered by ethical controversy. One would have had a reasonable expectation of success since Takahashi teaches human induced pluripotent stem cell have the same differentiation potential as human embryonic stem cells. One would have expected similar results since Lu teaches human embryonic stem cells and Takahashi teaches induced pluripotent cells behave the same way as human embryonic stem cells.
Therefore claim 187 is rendered obvious as claimed.
Therefore Applicant’s Invention is rendered obvious as claimed.
Claims 189 and 208 are rejected under pre-AIA 35 U.S.C. 103(a) as being unpatentable over Lu in view of Rajesh as recited in the rejection of claim 184 above, and further in view of Pick et al. (previously cited; Differentiation of Human Embryonic Stem Cells in Serum-Free Medium Reveals Distinct Roles for Bone Morphogenetic Protein 4, Vascular Endothelial Growth Factor, Stem Cell Factor, and Fibroblast Growth Factor 2 in Hematopoiesis. STEM CELLS 2007;25:2206–2214; Epub 2007 Jun 7).
Claim 184 is rejected on the grounds set forth above. The teachings of Lu are Rajesh are reiterated. Lu teaches culturing in a first serum free medium comprising 50 ng/ml BMP-4 and VEGF165.
Lu does not teach the first medium comprises bFGF (claim 189).
Lu does not teach using 25 ng/ml BMP-4.
Pick teaches a serum differentiation system to investigate the roles of four growth factors, bone morphogenetic protein 4 (BMP4), vascular endothelial growth factor (VEGF), stem cell factor (SCF), and basic fibroblast growth factor (FGF2), singly and in combination, on the generation of hematopoietic cells from human embryonic stem cells (HESCs) (Abstract). Pick teaches FGF can be combined with VEGF and BMP-4 (see Figure 3, BVF). Pick teaches FGF2 increases cell yield during human embryonic stem cell differentiation (see text of Figure 4). Pick teaches BMP-4 can be used at a concentration of 1-50 ng/ml and teaches it has a dose dependent effect on the percentage of hematopoietic cells obtained (see page 2208, left column second paragraph).
It would have been obvious to include bFGF in the first medium taught by Lu. One would have been motivated to do so since Pick teaches FGF2 can be combined with BMP4 and VEGF to increase cell yield. One would have had a reasonable expectation of success since Pick teaches FGF2 can be combined with BMP4 and VEGF. One would have expected similar results since both references are directed to human embryonic stem cell differentiation. Therefore claim 189 is rendered obvious.
It would have been obvious to use 25 ng/ml BMP-4 used in Lu’s method. Pick teaches BMP-4 can be used at a concentration of 1-50 ng/ml and teaches it has a dose dependent effect. The skilled artisan would do so to optimize the percentage of hematopoietic cells obtained for use in the in vivo and in vivo assays taught by Lu.
One would have had a reasonable expectation of success since Pick teaches the amount of BMP-4 can be optimized. One would have expected similar results since both references are directed to methods of differentiating human embryonic stem cels..
Therefore claim 208 is included in this rejection.
Therefore Applicant’s Invention is rendered obvious as claimed.
APPLICANT’S ARGUMENTS
The arguments made in the response filed on 10 April 2025 are acknowledged. The Applicant argues Pick, Rajesh and Crawford do not teach 25-100 ng/ml VEGF as now recited in the base claims.
EXAMINER’S RESPONSE
The rejection of the claims over Pick in view of Rajesh and Crawford have been withdrawn due to claim amendments. New grounds of rejection have been necessitated by claim amendment.
CONCLUSION
No Claims Are Allowed
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any 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.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to NATALIE MOSS whose telephone number is (571) 270-7439. The examiner can normally be reached on Monday-Friday, 8am-5pm EST.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Sharmila Landau can be reached on (571) 272-0614. The fax phone number for the organization where this application or proceeding is assigned is (571) 273-8300.
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/NATALIE M MOSS/ Examiner, Art Unit 1653
/SHARMILA G LANDAU/Supervisory Patent Examiner, Art Unit 1653