DETAILED ACTION
Claim Status
As of the Advisory Action mailed 5/5/2026, claims 1, 6-9, 16-17, and 19-27 were pending.
In Applicant's Response filed on 5/12/2026, claim 1 was amended.
As such, claims 1, 6-9, 16-17, and 19-27 are pending and have been examined herein.
Withdrawn Objections/Rejections
The objections and rejections presented herein represent the full set of objections and rejections currently pending in this application. Any objections or rejections not specifically reiterated are hereby withdrawn.
New Grounds of Rejections Necessitated by Amendments
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.
Claim(s) 1, 6-8, 16-17, 19-21, and 24-27 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sethu et al (US20120034695A1, 6/30/2011; Epub 2/9/2012) in view of Lim et al (US20170369904A1, 6/21/2017; published 12/28/2017; of record).
Sethu teaches a cell culture system and method of culturing (abstract). The system includes a cell culture chamber, membrane having cultured cells thereon, a pressure generator coupled to the culture platform (see claim 20 of Sethu). The method includes applying external pressure including compressing volume and increasing pressure in the tissue/cell culture chamber (see claim 25 of Sethu) and modulating the pressure inside the chamber (see claim 26 of Sethu). Figures 13A-F show waveform application of different pressure conditions. The reference teaches that the system can be used on both cardiogenic and non-cardiogenic cells including adult cardiomyocytes, skeletal myoblasts, smooth muscle cells, fibroblasts, endothelial progenitors, mesenchymal stem cells, hematopoietic stem cells, other marrow populations, resident myocardial progenitors, and embryonic stem cells (para 61). Regarding claim 1 in-part and claims 24-27, the reference does not explicitly teach fluctuation frequency of 12-100 times/minute, 13-18 times/minute, 40-80 times/minute, 13-15 times/minute, or 50 to 70 times per minute as instantly claimed. However, Sethu teaches that the applied pressure, percentage systolic/diastolic fraction and frequency can be manipulated to alter frequency and amplitude of pressure and flow waveforms. In addition to this chamber, tunable compliance and flow resistance elements upstream of the inlet (pulmonary) and downstream of the outlet of the cell culture channel (aortic/systemic) allow modulation of flow resistance and modification of shape and amplitude of attained pressure and flow profiles (para 109). Thus, it would have been a matter of routine optimization using standard laboratory techniques available at the time of filing to determine the appropriate amount of times to fluctuate the dynamic hydrostatic pressure as taught by the prior art. This reads on “a method of culturing stem cells, comprising culturing stem cells in an atmospheric environment comprising an additional pressure applied in addition to an atmospheric pressure, wherein said additional pressure changes by way of a sinusoidal or sinusoid-like periodic fluctuation . . . so that the additional pressure is a dynamic pressure; wherein the dynamic pressure is applied by connecting a chamber containing the stem cells and a pressure modulation device that is in gas communication with the chamber” as in instant claim 1 in-part; “wherein the stem cells are primary stem cells, passaged stem cells, or a combination thereof” as in instant claim 6; “wherein said stem cells are human stem cells” as in instant claim 7; “wherein said stem cells are mesenchymal stem cells” as in instant claim 8. In normal condition, the pressure applied is 120 mmHg at a rate of 80 beat per minute (bpm) and 40% of the cycle is diastolic phase (para 86). The pressure generator inserts air with high pressure (ex. 120 mmHg for the normal condition, or 180 mmHg for the hypertension case) to the pneumatic chamber via pneumatic inlet, pushing the circular post up to the original position (away from the first direction) which in turn, push the cell membrane back to its original position (away from the first direction) (para 86). Fig. 17B shows waveform pressure application above 50 mmHg and below 150 mmHg (see screenshot below) (overlaps with “wherein said additional pressure changes by way of a sinusoidal or sinusoidal-like periodic fluctuation within a range of 60 – 140 mmHg” as in instant claim 1 in-part; “wherein said additional pressure changes by way of the sinusoidal or sinusoid-like periodic fluctuation within a range of 70 - 120 mmHg” as in instant claim 16 and “wherein said additional pressure changes by way of the sinusoidal or sinusoid-like periodic fluctuation within a range of 75 - 115 mmHg” as in instant claim 17).
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The difference between the instant invention and Sethu is that it does not teach that the oxygen concentration is in a range of 2-8% (instant claim 1 in-part) or ranges between 3-7% (instant claims 19-21).
Lim teaches a method of increasing transfection efficiency and cellular reprogramming. The reference teaches that culturing a cell in hypoxic conditions and positive pressure (see, e.g., claims 17 or 18 of Lim). The reference teaches that moderate oxygen and moderate pressure levels can be used to efficiently propagate cells while maintaining, for example, pluripotency (para 94). The oxygen levels can vary from about 5% to about 15% (same para) (overlaps with “wherein said atmospheric environment comprises an oxygen concentration in a range of 2% - 8%” as in instant claim 1 in-part; “wherein said atmospheric environment comprises an oxygen concentration in a range of 3% - 7%” as in instant claim 19; “wherein said atmospheric environment comprises an oxygen concentration in a range of 4% - 7%” as in instant claim 20; “wherein said atmospheric environment comprises an oxygen concentration in a range of 5% - 7%” as in instant claim 21
Therefore, it would have been obvious prior to the effective filing date of the instantly claimed invention to culture stem cells under additional pressure as taught by Sethu, where the oxygen is between 5 and 15% as taught by Lim, to arrive at the instantly claimed invention. Lim shows that cells can be cultured in a combination of additional pressure and hypoxia between 5 and 15%. One of ordinary skill would have been motivated to modify the culture conditions of Sethu to include the hypoxic conditions of Lim with a reasonable expectation of advantageously propagating cells efficiently while maintaining pluripotency as taught by the prior art.
Claim(s) 9 and 23 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sethu and Lim as applied to claim 1, 6-8, 16-17, 19-21, and 24-27 above, and further in view of Thompson et al (US20100093083A1, 2/18/2008; published 4/15/2010).
The difference between the combined teachings and the invention as instantly claimed is that they do not teach that the stem cells are cultured between 36.5-37.5 degrees Celsius or that the culturing is at 37 degrees Celsius.
Thompson discusses culturing stem cells. The reference teaches that a number of parameters are important for maintaining and expanding stem cells in large-scale cultures, including temperature, oxygen concentration, pH, concentrations of nutrients (e.g. glucose) and waste products (e.g. lactate), and shear rate (para 32). In general, the temperature at which mammalian stem cells are cultured is about 37° C.±0.5° C., although the skilled person will appreciate that wider variations of temperature might be possible in some embodiments (same para). Typically, temperature is controlled by placing the culture vessels in temperature-controlled incubators or, in the case of some bioreactors, by direct monitoring and control of the temperature in the culture vessel. Temperature variations can be minimized by ensuring all media and culture components are brought to the required temperature before being added to the culture vessel (Same para). This reads on “wherein said culturing is conducted at a temperature in a range of 36.5 - 37.5 °C.” as in instant claim 9 and “wherein said culturing is conducted at a temperature of 37 °C” as in instant claim 23.
Therefore, it would have been obvious prior to the effective filing date of the instantly claimed invention to culture stem cells under additional pressure and hypoxia as taught by Sethu and Lim in combination, where the stem cells are cultured at 37 °C as taught by Thompson, to arrive at the instantly claimed invention. Thompson shows the temperature at which mammalian stem cells are cultured is about 37° C.±0.5° C., one of ordinary skill would have been motivated to modify the culture method of Sethu and Lim in combination to include culturing at 37 °C with a reasonable expectation of advantageously maintaining and expanding stem cells as taught by the prior art. "[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation." In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955) (see MPEP 2144.05(II)(A)). It also would have been a matter of routine optimization using standard laboratory techniques available at the time of filing to determine the appropriate temperature to culture the stem cells as taught by the prior art.
Claim(s) 22 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sethu and Lim as applied to claims 1, 6-8, 16-17, 19-21, and 24-27 above, and further in view of Dai et al (J Biomech. 2014 Mar 21; 47(5):966-72; of record).
The difference between the combined teachings and the invention as instantly claimed is that they do not teach that the stem cells are selected from the group consisting of human adipose mesenchymal stem cells, human endometrial mesenchymal stem cells, human hair follicle mesenchymal stem cells and human umbilical mesenchymal stem cells.
Dai teaches that dynamic compression and co-culture of adipose-derived mesenchymal stem cells with nucleus pulposus cells promotes proliferation and differentiation (title). The reference teaches that alginate beads were used to package the NPCs for separation from ASCs in T-25 culture flasks. NPCs were harvested from T-25 flask after 48-h culture and encapsulated in alginate beads in individual wells of six-well plates. The ten alginate beads encapsulating NPCs were co-incubated with ASCs (approximately 1:1 ratio) in a T-25 flask at 37 °C and 5% CO2 in DMEM/F12 with 10% heat-inactivated fetal bovine serum. Mechanical stimulation was achieved by a dynamic compression apparatus shown in Supplementary data Fig. 1. The flask pressure was controlled by adjusting the air pressure at both entrance and exit. An open pressure-adjusting system was created by placing the gas circulating system in the CO2 incubator, including a pressure pump, control device, flow valve and an incubator (Fig. 3). The pressure pump was set to provide intermittent dynamic hydrostatic pressure at 17 kPa under 220 V. The apparatus operated on alternating 12-h shifts of work and rest. It allowed 4 days of proliferation and 7 days of differentiation (“Co-culture and dynamic compression” para 1-2).
Therefore, it would have been obvious prior to the effective filing date of the instantly claimed invention to culture stem cells under additional pressure and hypoxia as taught by Sethu and Lim in combination, where the cells are adipose mesenchymal stem cells as taught by Dai, to arrive at the instantly claimed invention. As Dai shows that adipose mesenchymal stem cells can be cultured under dynamic pressure conditions, one of ordinary skill would have been motivated to simply substitute one known element (stem cell of Sethu and Lim) for another (adipose mesenchymal stem cell of Dai) to obtain the predictable result of advantageously promoting proliferation of the cells.
Response to Arguments
Applicant’s arguments regarding previously cited references Sugimoto and Guo have been fully considered but are moot as because the new ground of rejection does not rely on any reference applied in the prior rejection of record for any teaching or matter specifically challenged in the argument. Applicant has not provided any arguments challenging the specific teachings of cited reference Dai.
On p. 6-7 of Remarks, Applicant argues that the purportedly advantageous effects associated with the claimed combination of hypoxic conditions with dynamic pressure fluctuation shows increased expansion fold, increased colony-forming unit rate, enhanced differentiation potential, up-regulation of stemness-related genes, and downregulation of senescence-related genes.
In response, the examiner notes that, as per MPEP § 716.02, [a]ny differences between the claimed invention and the prior art may be expected to result in some differences in properties. The issue is whether the properties differ to such an extent that the difference is really unexpected. In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986). "A greater than expected result is an evidentiary factor pertinent to the legal conclusion of obviousness ... of the claims at issue." In re Corkill, 771 F.2d 1496, 226 USPQ 1005 (Fed. Cir. 1985). However, a greater than additive effect is not necessarily sufficient to overcome a prima facie case of obviousness because such an effect can either be expected or unexpected. Applicants must further show that the results were greater than those which would have been expected from the prior art to an unobvious extent, and that the results are of a significant, practical advantage. Ex parte The NutraSweet Co., 19 USPQ2d 1586 (Bd. Pat. App. & Inter. 1991) (see MPEP § 716.02(a)). Previously cited reference Dai shows that dynamic pressure conditions aids in proliferative capacity of mesenchymal stem cells. Previously cited Lim shows that a combination of positive pressure conditions and hypoxia efficiently propagate cells while maintaining pluripotency of the stem cells. Previously cited Guo (not utilized in any rejections above) shows that dynamic culture conditions (albeit application of mechanical pressure as opposed to air pressure) maintains high cell population. These teachings of the prior art, at the very least, show that Applicant’s purported advantageous effects were known in the prior art and would be expected by one of ordinary skill in the art prior to the effective filing date of the instant invention. Thus, Applicant’s arguments are not persuasive.
Conclusion
No claim is allowed.
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/G.R./Examiner, Art Unit 1632
/PETER PARAS JR/Supervisory Patent Examiner, Art Unit 1632
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