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 .
Status of Claims
Amendment filed on 01/15/2026 is acknowledged.
Claims 3-4, 13, 26, 28-56 remain cancelled. Claim 5 is now cancelled.
Claims 1-2, 6-7, 15, 19 and 21- 25 are amended. Claims 58-59 are new.
Claims 1-2, 6-12, 14-25, 27 and 57-59 are pending and being examined merits herein.
Priority
This instant application 17055276, filed on 11/13/2020, is a 371 of PCT/US2019/032155, filed on 05/14/2019, which claims domestic benefit of 62/671074, filed on 05/14/2018.
Information Disclosure Statement
The information disclosure statement (IDS), filed on 01/15/2026 is in compliance with the
provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the
Examiner.
Withdrawn Objections/Rejections
All previous claim Objection(s) / Rejection(s) as set forth in the previous Office action (mailed 09/30/2025) that are not repeated and/or maintained in the instant Office action are withdrawn, in light of applicant’s amendment and remark filed on 01/15/2026.
Claim Interpretation
Claims 1, 21, and 58-59 are interpreted below.
Claim 1 is interpreted as following:
A composition comprising an enriched population of extracellular vesicles, a hypertonic cell culture media bearing an osmolality between 382 and 1000 mOsm/kg.
Regarding phrases of “immersed in”, “wherein the hypertonic cell culture media can sustain growth of living cells in an incubated environment, wherein said enriched population of extracellular vesicles is produced by living cells incubated in the hypertonic cell culture media, wherein said enriched population of extracellular vesicles is at least 10% higher than a reference population of extracellular vesicles produced by the living cells incubated in an isotonic cell culture media bearing an osmolality between 250 and 320 mOsm/kg when incubation time are the same” are interpreted as properties or “intended use”, or “process of making” the composition, because they do not structurally contribute to the composition.
Claim 21 is interpreted as following:
A kit comprising:
living cells comprising an enriched population of extracellular vesicles;
a hypertonic cell culture media bearing an osmolality between 382 mOsm/kg and 20 Osm/kg (20,000 mOsm/kg).
The phrases of “wherein the hypertonic cell culture media can sustain growth of living cells in an incubated environment, wherein said enriched population of extracellular vesicles is produced by living cells incubated in the hypertonic cell culture media, wherein said enriched population of extracellular vesicles is at least 10% higher than a reference population of extracellular vesicles produced by the living cells incubated in an isotonic cell culture media bearing an osmolality between 250 and 320 mOsm/kg when incubation time are the same” are interpreted as properties or “intended use”, or “process of making” the composition, because they do not structurally contribute to the kit composition.
Claim 58 is interpreted as “intended use” or property of the composition of claim 1, because the “living cells remain viable …” does not structurally contribute to the composition.
Claim 59 is interpreted as the hypertonic cell culture media comprises additional sodium chloride, because the phrase “cell culture media spiked with” implies an action or process, and it does not contribute to the structure of the composition.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-2, 6, 14-16, 20-25 and 58 are rejected under 35 U.S.C. 103 as being unpatentable over Pollock et al. (Tissue Engineering: 2016, in record of 09/30/2025).
Pollock investigates multicomponent osmolyte solutions and their ability to preserve cell viability during cryopreservation to improve mesenchymal stem cells (MSCs) survival and discovers that maximum post-thaw recovery is observed in the multiple osmolyte solutions with incubation times of up to 2h before freezing (Abstract).
Pollock teaches SMC solution combining sucrose, mannitol and creatine with low osmolarities (<500 mOsm), and SGC solution containing sucrose, glycerol (considered small molecule, Pg. 1004, left bottom), and creatine with high osmolarity (<1200 mOsm) (the multicomponent solution containing nutrients for cell growth, thus corresponding to hypertonic cell culture media), and 10% DMSO solution with osmolarity of approx. 1400 mOsm (note: DMSO associates with poor post-thaw behavior and with dangerous systemic side effect, Pg. 999, Background), are used to test viability of mesenchymal stem cells (Pg. 1001, Influence of osmolarity and composition) (corresponding to hypertonic cell culture media in instant claims 1 and 21, and protein-free base media in instant claim 24). The stem cell population comprising extracellular vesicles in SGC solutions with osmolality ranging from 368-1187 mOsm recovers live cell fraction from 0.21 to 1.13, while in SMC solution with osmolality ranging from 303-552 mOsm resulting in live cell recovery from 0.09 to 0.54 (Appendix Table A1 and A2). Some solutions with high osmolarity showing very good live cell recovery, for instance, solution contains 225 mM sucrose and 684 mM glycerol (mOsm =1184) recovering live cell 0.94 fraction; adding 3 mM of creatine into this solution, osmolarity increases to 1187, while live cell recovery rate is 0.67. When SGC solution contains 75mM sucrose, 342 mM glycerol, and 25 mM creatine, live cell recovery faction is 1.13 at mOsm of 717 (Table A2), showing significantly higher values than the SMC solutions with lower osmolarity, e.g., mOsm at 301, live cell recovery fraction at 0.52; mOsm at 316, live cell recovery fraction at 0.31 or 0.33; mOsm at 303, cell recovery faction at 0.29 (Table A1). The live cell population which comprises extracellular vesicles is therefore enriched in SGC solution with high osmolarity levels, e.g., 1184, 1187, 717, compared to SMC solution as reference with low osmolarity, e.g., 301, 303, 316, for at least 15% higher population levels based on examples above (minimum increase: 0.67-0.52=0.15=15%, maximum increase: 1.13-0.29 =0.84=84%) (corresponding to instant claims 1, 16, and 21).
In light of claim interpretation, instant claims 1 and 21, the recitation of the phrase “the enriched population of extracellular vesicles is produced by living cells incubated in the hypertonic cell culture media” is interpreted as “intended use” or “process of making”. Prior art teaches the composition as instantly claimed; therefore the intended use would be necessarily capable of being obtained. If it is interpreted as “process of making”, the patentability is based on the product itself. MPEP indicates that the process of making is only relevant “if the process by which a product is made imparts ‘structural and functional differences’ distinguishing the claimed produce from the prior art”. See MPEP 2113.
Pollock teaches that human H9 ESC-derived mesenchymal stem cells (MSCs) (corresponding to instant claims 6 and 22), known as genetically modified cell type to have unique biological properties leading to development of MSC-based therapies for a wide range of diseases (Background, corresponding to instant claim 25), are cultured in alpha-MEM (Gibco) supplemented with nonessential amino acids and 10% FBS in a 37C incubator at 5% CO2, and media are changed every 3-4 days, and cells are used for experiments only between passages 8 and 12 (Pg. 1000, Cell culture) (corresponding to instant claim 23).
Pollock indicates that the cryoprotectant solutions, e.g., SGC solutions with high osmolality, are present at 2x their final concentrations (Pg. 1000, Cell freezing), resulting in double osmolality of corresponding solutions, for instance, mOsm at 1187 of specific SGC solution would present 2374 mOsm (=2.374 Osm/kg), and are added an equal volume to the cell-containing vials to reach final concentration (corresponding to instant claim 2), then the cell suspensions are proceeded to incubation at room temperature for up to 4h before freezing according to designated protocol (Pg. 1000, Cell freezing) (corresponding to instant claim 14 ). In Raman measurement of frozen MSC cells, Pollock specifies that MSC cells are washed with DPBS solution before being suspended in experimental solutions, and then the cell suspension is placed on an aluminum sheet, covered with a piece of mica (Ted Pella, Redding, CA) (implying immobilization or encapsulation in another material) and sealed with Kapton tape (DuPont, Wilmington, DE), to prevent evaporation/sublimation during each experiment, and then cooled and frozen (Pg. 1001, left middle)(corresponding to instant claim 14) , indicating that the composition is in a form of a patch (corresponding to instant claim 20). Therefore, in summary, Pollock teaches a kit suspending the cells in the solution with defined osmolality, or keeping the cells with mica in a patch form, wherein comprising the living stem cells comprising extracellular vesicles and a solution bearing an osmolality possibly higher than 382 mOsm/kg to 1187 mOsm/kg and comprising an in vitro cell culture media, wherein the cell or extracellular vesicles population at 15% higher or more compared to a reference population in a solution with osmolality around 300 mOsm/kg (corresponding to instant claim 21).
Pollock teaches that multiple families of cryoprotectants, such as amino acids, trehalose and proline, or trehalose and glycerol, exhibit improved cellular survival compared with either cryopreservative alone, suggesting that additive or synergistic stabilizing effects are possible when multiple cryopreservatives are used (Pg. 1000, middle paragraph), and Pollock indicates that solutions can contain multiple cryoprotectants, including sugars (as carbohydrate), sugar alcohols, and small molecule additives (Abstract) (corresponding to instant claim 15).
Pollock investigates cell viability incubated in the multicomponent cell culture SGC and SMC at room temperature (overlapping with temperature range between 4 and 45 C in instant claim 58) for up to 4h, and Pollock indicates that longer incubation times (4h) decreases the viability (Pg. 1001, right middle), suggesting that longer incubation time is possible yet with tradeoff of decreased viability. For the purpose of preserving cells is to enable transportation of cells from site of manufacture to the site of administration and coordination of the therapy with patient availability (e.g., Pg. 999, right side), longer time duration, e.g., 12 hr, of keeping the cells in such media can be inevitable. As evidenced by instant specification that cell viability can be any amount between 50-100% (Pg. 8, line 8), even if decreased viability to 50% is acceptable. Further, in light of claim interpretation, instant claim 58 is interpreted as “intended use” or property of the composition, which has been taught by prior art, therefore, the property or “intended use” would necessarily present or capable of being achieved.
Pollock does not explicitly teach that the hypertonic cell culture media (i.e., SGC with high osmolality at 717, 1184, or 1187 mOsm/kg) can be used for enriched extracellular vesicles, however, living cells comprise extracellular vesicles. Therefore, an enriched living cell population would obviously include an enriched extracellular vesicle population. Moreover, Pollock suggests that these solution compositions being successful could be the result of their interactions to stabilize proteins within the cell and in the membrane and different combinations of osmolytes stabilizes the cell membrane (Pg. 1005, left bottom and right top paragraph). Pollock further teaches that larger sugar osmolytes provide better stabilization due to their increased polar contact area, and sugars provide stabilization by replacing water surrounding membranes during dehydration, perhaps due to insertion between the phospholipid heads of the lipid membrane, creating more space than monosaccharides for additional binding, and this additional binding of sugars to membranes can increase the rigidity of the membrane, which provides greater resistance to disruption (Pg. 1005, right bottom paragraph). Since extracellular vesicles are membrane-rich organelles in cells, a person with ordinary skills in the art would have motivation to use these components/solutions to enhance the membrane stability and integrity during extracellular vesicles preparation.
A person with ordinary skill in the art would be motivated to take the opportunity to implement and optimize Pollock’s teaching to reach current invention. Because Pollock specifies that ability to preserve cells enables transportation of the cells from the site of manufacture to the site of administration and coordination of the therapy with patient availability, and conventional methods of using DMSO constitute dangerous systemic side effects and poor post-thaw behavior (Pg. 999, Background), it provides artisans in the field good reasons for the development of non-DMSO in-vitro cell culture media in need for preservation of living cells comprising extracellular vesicles. Since Pollock establishes a composition with non-DMSO in vitro cell culture media constituting relatively high osmolality that can greatly enhance living stem cell population viability during incubation, freezing, transportation, thawing, and Pollock teaches that the solutions with high osmolality containing sugar potentially enable cell membrane stabilization as well as membrane protein or in-cell protein stabilization, which logically improve stabilization and enrichment of extracellular vesicles along with living cells. This renders obviousness as “use of known technique to improve similar devices (methods, or products) in the same way” or as “applying a known technique to a known device (method, or product) ready for improvement to yield predictable results”. See MPEP §2143. (I)(C) and (I)(D). Moreover, It is prima facie obvious to select a known material for incorporation into a composition, based on its recognized suitability for its intended use (MPEP §2144.07). See Sinclair & Carroll Co. v. Interchemical Corp., 325 U.S. 327, 65 USPQ 297 (1945).
Generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature is critical. See In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955). MPEP §2144.05(I) states that “A prima facie case of obviousness typically exists when the ranges of a claimed composition overlap the ranges disclosed in the prior art.” See In re Peterson, 315 F.3d 1325, 1329 (Fed. Cir. 2003). The osmolarities between 382-1000 mOsm/kg of the target culture solution or the reference solution of 250-320 mOsm/kg in instant claim 1, or 1-20 Osm/kg in instant claim 2, or 382 mOsm/kg-20 Osm/kg of the target solution and 250-320 mOsm/kg reference solution, overlap with those taught by Pollock. Even though Pollock does not explicitly teach that in vitro cell culture media solution, i.e., SGC with high osmolality (e.g., mOsm 717, 1184, 1187) can be used for enriched extracellular vesicles since living cells comprise extracellular vesicles. The osmolarities used for enriching living cell population in prior art would obviously enrich extracellular vesicle population, and therefore, Pollock’s teaching would have provided a reasonable expectation of success for an artisan in the field to implement these high osmolality solutions and optimize them in extracellular vesicle production and enrichment. MPEP §2144.05 (II) states that “[i]t would have been prima facie obvious for one of ordinary skill in the art to optimize additive amount through nothing more than “routine experimentation,” because of a reasonable expectation of success resulting from the optimization for desirable features of intended use of the composition. See Peterson, 315 F.3d at 1330, 65 USPQ2d at 1382; In re Hoeschele, 406 F.2d 1403, 160 USPQ 809 (CCPA 1969).
Claims 1-2, 6-11, 14-17, 19-25, 27 and 57-58 are rejected under 35 U.S.C. 103 as being unpatentable over Pollock et al. (Tissue Engineering: 2016, in record of 09/30/2025) as applied to claims 1-2, 6, 14-16, 20-25 and 58 above, further in view of Hurwitz et al. (Journal of Virology, March 2018, in record of 09/30/2025).
As discussed in great detail above, Pollock teaches a composition or kit comprising multicomponent hypertonic cell culture media, e.g., SGC solution, with high osmolality up to 1187 mOsm/kg to improve living mesenchymal stem cells (MSCs) viability during incubation, freezing, transportation, and thawing, with significant increase as high as 84% of living cell recovery compared to reference low osmolality culture solution, e.g., about 300 mOsm/kg.
Pollock does not teach the cells are immortalized primary cells as recited in instant claim 7, enriched population comprising a cargo as recited in instant claim 8, cargo being endogenous as recited in instant claim 9, cargo being CD9, CD63, CD81 and others as recited in instant claim 10, enrich population of extracellular vesicles or cargo constituting those functions as recited in instant claim 11, vesicles having particle diameter between 50-1000 nm as recited in instant claim 17, cargo comprising those species as recited in instant claim 19, extracellular vesicles being modified to change, add, or remove cargo from vesicles as recited in instant claim 27, or cargo being exogenous as recited in instant claim 57.
Hurwitz throughout the reference teaches tetraspanin protein CD63 (corresponding to instant claim 15) as a key factor in extracellular vesicle production and endosomal cargo sorting, e.g., latent membrane protein 1 (LMP1) of EBV (Epstein-Barr virus, a human gamma herpesvirus) packaging (e.g., Abstract). Hurwitz presents in materials and methods growing HEK293 cells, as immortalized primary cells known in field (corresponding to instant claims 5-7 and 22), in DMEM cell culture supplemented with 10% FBS, 2 mM glutamine, and 100 IU penicillin-streptomycin and 100 ug/ml : 0.25 ug/ml antibiotic-antimycotic, and HK1 cells in RPMI 1640 cell culture medium with equivalent supplements. CD63 CRISPR (CRISPR known in art as genome editing tool) is introduced into the cells as an endogenous cargo. Hurwitz specifies that extracellular vesicles are enriched by ultracentrifugation at 1000,000 x g for 20 h and then filtered through a 0.2-um filter (Pg. 17, Materials and methods, Cell culture). LMP1 expression in HEK293 cells is carried out by doxycycline induction to cells for 24 h before cell lysate and EV collection, in other words, LMP1 is expressed via transduction, or transitory expression on EV. Therefore, the enriched extracellular vesicles of Hurwitz comprise endogenous cargo CD63 through gene modification (corresponding to instant claims 8-10, 22, and 25), and an exogenous cargo LMP1 (corresponding to instant claim 57), demonstrating extracellular vesicles have been modified to change, add cargos (corresponding to instant claim 27). Hurwitz describes cargos, e.g., GFP, GFP-LMP1, CD63, are constructed into plasmids which contain nucleic acid sequences and eventually express as cargo proteins via transformation into genome as endogenous cargos or via transduction for transient expression as exogenous cargos (e.g., pg. 17, Materials and Methods, DNA constructs) (corresponding to instant claim 19). Hurwitz indicates that cells with expression plasmids, e.g., GFP LMP1, packaging plasmid to produce retroviral particles for transduction is harvested at 48, 72, and 96 h post transfection (Pg. 17, Materials and methods, Retrovirus production), indicating culture time can be more than 3 days in the medium without hypertonic solution (corresponding to instant claim 23).
Hurwitz concludes that CD63 is required for proper protein packaging into small extracellular vesicles, with 331 unique proteins out of 1,200 being specific expressed in control compared to CD63 CRISPR cell-derived extracellular vesicles, with many involved in protein translation and vesicle transport, key proteins in signal transduction pathways (Fig. 1D; Table S1, Pg. 4, Results, 1st paragraph), and CD63 negatively regulates LMP1-mediated cellular proliferation, which may positively impact on Epstein-Barr virus associated tumorigenesis (Pg. 4, Results, 3rd paragraph). Therefore, cargos on enriched extracellular vesicles regulate protein secretion, cellular proliferation, and signal transduction (corresponding to instant claim 11).
Hurwitz states that electron microscopy confirms small vesicle-shaped particles ranging from 50 to 250 nm in size (Fig. 1B), and nanoparticle tracking analysis shows the mode and mean sizes of extracellular vesicles to be between 150 to 250 nm (Fig. 1C) (Pg. 4, Results, 1st paragraph) (falling within vesicle diameter between 50 and 1000 nm in instant claim 17). Hurwitz indicates that the vesicles may comprise other exosome subpopulations or nonexosomal vesicles (Pg. 4, Results, 2nd paragraph) (corresponding to instant claim 16).
It would have been prima facie obvious for a person with ordinary skills in the art to incorporate Hurwitz teaching of cargos for extracellular vesicles into the Pollock in vitro culture media solution to reach current invention. Substituting cells with cargos from Hurwitz into Pollock’s composition with high osmolality solution would retain the advantageous features from both prior art’s teachings, and would have maximized the benefit of stabilizing and enriching extracellular vesicle population, because Hurwitz demonstrates that cargos are important factors to stabilize and impact extracellular vesicles production and protein secretion and packaging, while Pollock provides the culture medium solution with high osmolality for enrichment and membrane integrity of extracellular vesicles. Combining them provides reasonable expectation of success in living cell population enrichment as well as extracellular vesicles. Therefore, the claimed invention is a simple combination of reagents known to be obvious materials that all already taught in prior art and discussed above. The idea for combining them flows logically from them having been individually taught in the prior art. In re Kerkhoven, 626 F.2d 846, 850, 205 USPQ 1069, 1072 (CCPA 1980). Consequently, it is prima facie obvious to implement the vesicle diameters with Hurwitz teaching of cargos for extracellular vesicles from immortalized primary cells into Pollock’s solution. Generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature is critical. See In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955). MPEP §2144.05(I) states that “A prima facie case of obviousness typically exists when the ranges of a claimed composition overlap the ranges disclosed in the prior art.” See In re Peterson, 315 F.3d 1325, 1329 (Fed. Cir. 2003). The vesicle diameters overlap with those taught by Hurwitz. More importantly, Hurwitz indicates that no difference in size of extracellular vesicles secreted from control or treatment groups (Pg. 11, bottom paragraph), suggesting these are ordinary extracellular vesicle sizes.
Claims 1-2, 6-12, 14-17, 19-25, 27 and 57-58 are rejected under 35 U.S.C. 103 as being unpatentable over Pollock et al. (Tissue Engineering: 2016, in record of 09/30/2025) and Hurwitz et al. (Journal of Virology, March 2018, in record of 09/30/2025) as applied to claims 1-2, 6-11, 14-17, 19-25, 27 and 57-58 above, further in view of Than et al. (International Journal of Molecular Sciences, 2017, 18, 956, 20 pages, IDS of 06/29/2022).
Pollock and Hurwitz combined teaching teaches compositions comprising enriched extracellular vesicles in high osmolality (up to 1187 mOsm/kg) in vitro culture solution with population higher than reference up to 84% in specific examples, and enriched extracellular vesicles can comprise cargos which can regulate protein secretion, signal transduction, cellular proliferation, and impact viral protein packaging on vesicles, as discussed above in great detail and incorporated herein.
Pollock and Hurwitz does not teach cargo is angiopoietin (Ang-1) as recited in instant claim 12.
Than teaches extracellular membrane vesicles associate with cutaneous wound healing (Title) and introduces different vesicle types including apoptotic bodies, microvesicles (ectosomes), exosomes with their specifics including diameter ranges, e.g., microvesicles between 100-1000 nm (Pg. 2-5). Throughout the reference Than teaches extracellular vesicles can carry DNA, small RNAs, proteins and membrane lipids, capable of regulating many biological processes, such as cancer progression, the immune response, cell proliferation, cell migration and blood vessel tube formation (e.g., Abstract) (corresponding to instant claims, e.g., claims 11, 15). Than teaches that internalization of extracellular vesicles by dendritic cells requires participation of surface molecules such as externalized PS, CD11a, CD54, CD9 and CD81 (Pg. 6, 3rd paragraph) (corresponding to instant claims 8-10). Than states that new blood vessel formation is critical for wound healing in order to supply nutrients and oxygen to newly formed tissues and that the formation of new blood vessels requires the proliferation of endothelial cells, as well as the interaction between endothelial cells, angiogenic factors; and extracellular vesicles contribute to the regulation of vessel formation by enhancing the expression of pro-angiogenic factors (Pg. 10, 5.4. Angiogenesis, 1st paragraph). Than then points out that various critical pro-angiogenic genes, including IL-6, IL-8, angiopoietin-1 (corresponding to instant claim 12), E-selectin, and fibroblast growth factor 2 are up-regulated after platelet-rich plasma and endothelial progenitor cell-derived exosome stimulation (Pg. 10, 5.4. Angiogenesis, 2nd paragraph).
It would have been prima facie obvious for a person with ordinary skills in the art to incorporate Than’s teaching of angiopoietin-1 as a cargo into the composition taught by Pollock and Hurwitz, because Than indicates how important these angiogenic factors impact the new blood vessel formation, and extracellular vesicles contribute to the blood vessel formation and can enhance expression of pro-angiogenic factors like angiopoietin-1. It is prima facie obvious to select a known material for incorporation into a composition, based on its recognized suitability for its intended use (MPEP §2144.07). See Sinclair & Carroll Co. v. Interchemical Corp., 325 U.S. 327, 65 USPQ 297 (1945).
Claims 1-2, 6, 14-16, 18, 20-25 and 58 are rejected under 35 U.S.C. 103 as being unpatentable over Pollock et al. (Tissue Engineering: 2016, in record of 09/30/2025) as applied to 1-2, 6, 14-16, 20-25 and 58 above, further in view of Deregibus et al. (Int. J. Mol. Med, 2016, in record of 04/11/2024).
As discussed in great detail above, Pollock teaches a composition comprising in vitro culture medium solution, e.g., SGC solution, with high osmolality up to 1187 mOsm/kg to improve living mesenchymal stem cells (MSCs) viability during incubation, freezing, transportation, and thawing, with significant increase as high as 84% of living cell recovery compared to reference low osmolality culture solution, e.g., about 300 mOsm/kg, resulting in enriched extracellular vesicles in the solution.
Pollock does not teach extracellular vesicles bear negative surface charge, or a negative zeta potential as recited in instant claim 18.
Deregibus teaches that extracellular vesicles have a negative charge based upon zeta-potential analysis and the negative charge allows interaction with positively charged molecules such as protamine, while protamine can induce extracellular vesicles precipitation from serum and saliva and from cell culture media without the need for ultracentrifugation, and resuspension of extracellular vesicles is facilitated when protamine precipitation is performed, with higher efficiency and enrichment of exosomes obtainable than ultracentrifugation (Abstract).
It would have been prima facie obvious for one with ordinary skills in the art prior to filing date to incorporate Deregibus’ teaching of extracellular vesicles being negative charged on surface or having zeta-potential into the composition comprising extracellular vesicles taught by Pollock. Since Deregibus teaches that the feature of extracellular vesicles being negatively charged is advantageous regarding potential improvement proficiency and facilitation in extracellular vesicles preparation, an artisan would have the motivation to take advantage of what is taught by Deregibus for reasonable expectation of success in extracellular vesicles production, bearing the same intention to potentially enrich target population with the culture medium constituting high osmolality taught by Pollock. It is prima facie obvious to select a known material for incorporation into a composition, based on its recognized suitability for its intended use (MPEP §2144.07). See Sinclair & Carroll Co. v. Interchemical Corp., 325 U.S. 327, 65 USPQ 297 (1945).
Claims 1-2, 6, 14-16, 20-25 and 58-59 are rejected under 35 U.S.C. 103 as being unpatentable over Pollock et al. (Tissue Engineering: 2016, in record of 09/30/2025) as applied to claims 1-2, 6, 14-16, 20-25 and 58 above, further in view of Blak et al. (US20130224857, 08/29/2013, IDS of 06/29/2022).
Pollock teaches a composition or kit comprising multicomponent hypertonic cell culture media, e.g., SGC solution, with high osmolality up to 1187 mOsm/kg to improve living mesenchymal stem cells (MSCs) viability during incubation, freezing, transportation, and thawing, with significant increase as high as 84% of living cell recovery compared to reference low osmolality culture solution, e.g., about 300 mOsm/kg, as discussed and applied to claims 1-2, 6, 14-16, 20-25 and 58 above in great detail and incorporated herein.
Pollock does not teach the hypertonic cell culture media comprises additional sodium chloride as in instant claim 59.
Blak throughout the reference teaches methods for generating germ layers from stem calls comprising the stem cells in a culture media having osmolality ranges that promote the generation of specific germ layer progenitor cells (e.g., Abstract).
Blak teaches that for endoderm differentiation, the embodiment comprises pluripotent stem cells are cultured in a media with lower osmolality such as 260-360 mOsm/kg [0039], and of higher osmolality such as 330-550 mOsm/kg (e.g., [0039]), or 320-550 mOsm/kg (e.g., Claims 14 and 17; [0051]), or 350-450 mOsm/kg (e.g., Claim 18; [0052]), or 290-550 mOsm/kg (Claim 13), for generating a population of enriched endodermal progenitor cells (e.g., [0051], [0052]), the endodermal progenitor cell population would inevitably contain enriched extracellular vesicles. Blak teaches that osmolality can be adjusted using addition of compounds, such as sodium chloride to make greater than 330 mOsm/kg (e.g., [0057]; [0382]; Claim 33), corresponding to instant claim 59.
It would be prima facie obvious for a person with ordinary skills in the art to incorporate Blak’s teaching to add sodium chloride as a component to the culture media to increase osmolality into the composition taught by Pollock to arrive at current invention. Because both Blak and Pollock are using higher osmolality culture media for enriched cell population (which results in enriched extracellular vesicle population) with good cell viability, to add the simple compound sodium chloride to enable osmolality increase would have motivated artisans to experiment and it would have provided reasonable expectation of success for an increased osmolality. It is prima facie obvious to select a known material for incorporation into a composition, based on its recognized suitability for its intended use (MPEP §2144.07). See Sinclair & Carroll Co. v. Interchemical Corp., 325 U.S. 327, 65 USPQ 297 (1945).
Response to Arguments
Applicant's arguments filed 01/15/2026 have been fully considered.
Applicant asserts that Pollock describes removing cell culture media before placing the cells in the high osmolality SGC solution for cryopreservation for the intention of stopping cell growth and metabolism to maintain cell health, while the instant claims emphasize that the claimed enriched extracellular vesicle population is formed by the incubation of the living cells in this hypertonic cell culture media.
In response, present office action has adjusted art rejections in light of claim amendments. Because Pollock teaches the high osmolality SGC solution contains multicomponent and nutrients, and keeps cells in the cryopreservation solution for preservation or transportation, therefore, it is practically a cell culture media with high osmolality. Pollock presents the enriched population resulting from the incubation of the cells in the hypertonic SGC with an approx. 84% living cell recovery, the enrichment is thus formed by the incubation of the living cells in this hypertonic cell culture media, because it is obvious that without this hypertonic cell culture incubation, or else the enrichment would not have occurred. In other words, the hypertonic culture media in Pollock is keeping cells alive and maintaining cell health, and therefore it is inevitably a cell culture media.
Further, in light of claim interpretation as presented above, as presented in office action, the enriched extracellular vesicle population being formed by the incubation of the living cells in this hypertonic cell culture in instant claims 1 and 21 is interpreted as “intended use” or “process of making” of the composition. Pollock teaches the composition, therefore, the “intended use” of the composition can be necessarily obtained. If it is interpreted as “process of making”, the patentability is based on the product itself. MPEP indicates that the process of making is only relevant “if the process by which a product is made imparts ‘structural and functional differences’ distinguishing the claimed produce from the prior art”. See MPEP 2113.
In conclusion, the argument is not persuasive. Prior art teaches the current invention.
Conclusion
No claim is allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/DX.Z./Examiner, Art Unit 1616
/SUE X LIU/Supervisory Patent Examiner, Art Unit 1616