DETAILED ACTION
Claims 1-10 are pending.
Claim Objections
Claim 1 is objected to because of the following informalities: The claim recites an abbreviation and/or acronym of “PI working solution”, which should be spelled out at their first usage followed by the abbreviation/acronym in parenthesis to recite “propidium iodide (PI) working solution”. Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim 1 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 1 recites “no time sequence restriction exists between steps 1) - 3) and steps 4) - 5)”. It is unknown and unclear what “time sequence restriction” is being referred to. Does “time sequence” refer to the order in which the steps occur? Or does “no time sequence restriction” mean that there is no time delay between steps? Further, “between steps 1) - 3) and steps 4) - 5)”, is unclear. If the “no time sequence restriction” is referring to there being no time delay between steps, is it directed to the step between step 3 and 4, or is it referring to all of the steps? The metes and bounds of “no time sequence restriction exists between steps 1) - 3) and steps 4) - 5)” is unknown and one would not know what is being referred to.
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.
1. Claims 1, 3-5, and 7-10 are rejected under 35 U.S.C. 103 as being unpatentable over Gang et al., “In vitro mesengenic potential of human umbilical cord blood-derived mesenchymal stem cells” Biochemical and Biophysical Research Communications. Vol. 321, Issue I. Pages 102-108 doi:10.1016/j.bbrc.2004.06.111, in view of Todtenhaupt et al., “A robust and standardized method to isolate and expand mesenchymal stromal cells from human umbilical cord” Cytotherapy. Vol. 25, Issue 10. Pages 1057-1068 https://doi.org/10.1016/j.jcyt.2023.07.004.
Gang teaches a determination method for purity and population doubling time of human umbilical cord mesenchymal stem cells (hUC-MSCs), comprising the following steps:
1) mixing cultured hUC-MSCs with a buffer solution to obtain cell suspension (see entire document, see page 103 under “isolation and culture of UCB-derived adherent cells” and “Cell cycle analysis and proliferation studies”);
2) mixing and incubating the cell suspension obtained in step 1) with a fluorescein-labeled antibody to obtain an incubated substance (see figure 2 with legend, see page 103 under “Immunophenotyping of UCB-derived adherent cells by flow cytometry”);
3) centrifuging the incubated substance obtained in step 2), cleaning the precipitate obtained and then mixing with the buffer solution to obtain a substance to be tested, and the substance to be tested is used to determine the positive expression ratio of cells (see page 103 under “Chondrogenic differentiation and evaluation”, see page 103-104 under “Total RNA extractiuon and RT-PCR”);
5) centrifuging the immobilized substance obtained in step 4), mixing, incubating and centrifuging the precipitate obtained with a PI working solution (see page 103 under “Cell cycle analysis and proliferation studies”), cleaning the precipitate obtained, then mixing with the buffer solution (see page 103 under “Cell cycle analysis and proliferation studies”), and analyzing population doubling time using Multicycle for DNA software; no time sequence restriction exists between steps 1) - 3) and steps 4) - 5) (see page 103 under “Cell cycle analysis and proliferation studies”, see page 104 under “Morphological and proliferation characteristics of human UCB-derived adherent cells”) (instant claim 1). Gang teaches the antibody of step 2 being anti-human CD14 and an FITC-labeled or PE-labeled antibody is used (see page 103 under “Immunophenotyping of UCB0derived adherent cells by flow cytometry”) (instant claim 4). Gang teaches the conditions of incubation in step 2) include the temperature being 4ºC, the time being 30 minutes, and the incubation being conducted away from light (see page 103 under “Cell cycle analysis and proliferation studies” and “Immunophenotyping of UCB-derived adherent cells by flow cytometry”) (instant claim 5). Gang teaches the buffer solution being PBS (see page 103) (instant claim 10). Gang teaches the PI working solution being 50 µg/ml, the temperature being 20-25ºC, the time being 30 minutes and incubation is conducted away from light (see page 103 under “Cell cycle analysis and proliferation studies”. It is widely known that room temperature is around 20-22ºC, thus meeting the limitation of the temperature being 20-25ºC) (instant claim 9).
While Gang does not explicitly teach the ratio of buffer to hUC-MSC, nor the ratio of cell suspension to fluorescein-labeled antibody, nor the ratio number of the hUC-MSCs to volume of the buffer solution to the volume of the anhydrous ethanol solution being 1 x 105:1 ml: 3 ml and the volume percent of anhydrous ethanol being 75%, it would have been obvious to one of ordinary skill in the art to routinely optimize the ratios of buffer to hUC-MSC, the ratio of cell suspension to labeled antibody, and the ratio of the hUC-MSC to buffer solution to anhydrous ethanol. Thus, absent some demonstration of unexpected results from the claimed parameters, it would have been prima facia obvious to have arrived at the claimed ratios (hUC-MSCs to volume of buffer being 1x105:0.5 ml and cell suspension to fluorescein-labeled antibody being 100:1, the hUC-MSCs: buffer: anhydrous ethanol being 1 x 105:1 ml: 3 ml) as an obvious matter of routine optimization, namely performing routine experimentation to determine the optimum and workable ratios. As noted in In re Aller, 105 USPQ 233 at 235, More particularly, where 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 (instant claims 3-4 and 7).
Gang does not teach mixing the cultured hUC-MSCs in a buffer solution with an anhydrous ethanol solution, nor does Gang teach the temperature being 4ºC and the time is 12-24 hours.
Todtenhaupt teaches the use of a buffer solution (see entire document, see page 1059 “Before staining, the cells were washed twice with PBS and once with milliQ”) and anhydrous ethanol with hUC-SMCs (see page 1059) (instant claim 1). Todtenhaupt teaches the temperature being 4ºC and the time is 12-24 hours (see page 1061 under “Time limits for UC collection and storage”) (instant claim 8).
It would have been obvious to one of ordinary skill in the art at the time of the instant application to combine the methods of isolation, culturing, and analysis of hUC-MSCs taught by Gang, with the use of anhydrous ethanol to fix and stain hUC-MSCs taught by Todtenhaupt. Todetenhaupt teaches the use of anhydrous ethanol to dehydrate the slides, which results in the fixing of hUC-MSCs to the slide (see page 1059). It is widely known in the art that the use of anhydrous ethanol will fix/immobilize the hUC-MSCs to the slide. The artisan would have reasonable expectation of success based on the cumulative disclosure of these prior art references at the time the instant application was filed.
2. Claims 2 and 6 are rejected under 35 U.S.C. 103 as being unpatentable over Gang and Todtenhaupt as applied to claims 1, 3-5, and 7-10, in view of Guo et al., (US11091739B2) (effectively filed on 12/11/2015).
The teachings of Gang and Todtenhaupt as they pertain to claims 1, 3-5, and 7-10 are discussed in the 35 USC 103 rejection above.
Specifically, Gang teaches centrifuging for 5 minutes (see page 103 under “Chondrogenic differentiation and evaluation”) (instant claim 6).
Gang does not teach the passage number being 3rd to 6th generation, nor does Gang teach the revolving speed being 1200 rpm.
Gao teaches the hUC-MSC being 3rd generation (see entire document, see column 6 lines 9-11, see column 8 lines 25-26, see column 10 lines 20-26) (instant claim 2). Gao teaches the temperature being 4ºC and 1200 rpm (see column 9 lines 40-59) (instant claim 6).
It would have been obvious to one of ordinary skill in the art at the time of the instant application to combine the methods of isolation, culturing, and analysis of hUC-MSCs taught by Gang, with the use of anhydrous ethanol to fix and stain hUC-MSCs taught by Todtenhaupt, with the methods of hUC-MSC culturing taught by Gao. Gao teaches that the use of cultured hUC-MSC can maintain good proliferation and multi-directional differentiation potential during long-term culture (see column 4 lines 27-31). The artisan would have reasonable expectation of success based on the cumulative disclosure of these prior art references at the time the instant application was filed.
Conclusion
No claim is allowed.
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/MCKENZIE A DUNN/ Examiner, Art Unit 1678
/GREGORY S EMCH/ Supervisory Patent Examiner, Art Unit 1678
