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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 03 September 2025 has been entered.
DETAILED OFFICE ACTION
This Office Action is in response to the papers filed on 03 September 2025.
CLAIMS UNDER EXAMINATION
Claims 6, 10, 30-32, 38, 40 and 45-52 have been examined on their merits.
PRIORITY
A certified translation of the foreign application filed on 07/16/2014 has not been provided.
WITHDRAWN REJECTIONS
The previous rejections have been withdrawn in light of the Declaration filed on 03 September 2025.
REJECTIONS
New grounds of rejection are set forth below.
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 6, 10, 30-31, 38, 40 and 45-52 are rejected under 35 U.S.C. 103 as being unpatentable over Hattori et al. (previously cited; Method For Purifying Cardiomyocytes or Programmed Cardiomyocytes Derived From Stem Cells or Fetuses. US20090275132) in view of Mitcheson et al. (previously cited; Cultured adult cardiac myocytes: Future applications, culture methods, morphological and electrophysiological properties, Cardiovascular Research: 39 (1998) 280-300) and Sheen et al. (Defective Regulation of Autophagy upon Leucine Deprivation Reveals a Targetable Liability of Human Melanoma Cells In Vitro and In Vivo
Cancer Cell. 2011: 19, 613-628).
Hattori et al. teach a method of purifying cardiomyocytes from a cell mixture at a high degree of purification ([0017]). Hattori teaches the presence of “non-cardiomyocytes” that are selected to undergo cell death ([0019]). Embryonic stem cells (pluripotent stem cells) are differentiated to produce cardiomyocytes ([0052]).
Hattori studied the “compositions of various kinds of culture medium in order to construct a system for efficiently producing cardiomyocytes” ([0018]). Hattori teaches changing the composition of components of culture medium to efficiently select cardiomyocytes using various conditions ([0018] [0063]). Hattori prepares medium with the desired components by mixing commercially available mediums (see [0080]), diluting commercial mediums (see [0081]) and supplementing commercial mediums with the desired components (see [0081]).
The art teaches selection using a low glucose (sugar) condition and a low nutrient condition ([0060]). The art teaches one or more condition can be used ([0026]).
Regarding the low sugar condition: Hattori teaches selection of cardiomyocytes by culturing under “sugar-free” conditions ([0037]). When the culture medium is deprived of sugar, non-cardiomyocytes undergo cell death ([0071]). Hattori teaches it is effective to supplement the culture medium with a lactic acid (lactate, 0.1-5 mM) or a pyruvic acid (pyruvate) (0.5-5 mM) in place of the sugars ([0071] [0072]).
Hattori teaches the following:
differentiation from pluripotent stem cells to cardiomyocytes;
a mixture of cardiomyocytes and undifferentiated pluripotent stem cells;
culturing under sugar free conditions; and
supplementing with lactic acid or pyruvic acid.
Hattori teaches changing compositions and components of various culture mediums to produce the desired conditions.
Hattori teaches low-nutrient conditions.
The reference does not explicitly teach culturing in a medium that is also free of glutamine.
Mitcheson teaches a method of culturing cardiomyocytes (see title; Abstract). In the section titled “Medium changes”, Mitcheson teaches glutamine free conditions do not support cell division, avoiding the problems of contamination of myocyte cultures by rapidly dividing non-myocytes” (page 284, right column, third paragraph). Therefore Mitcheson teaches culturing in cell culture medium that does not contain glutamine to prevent contamination of myocyte culture with dividing non-myocytes.
Sheen teaches an amino acid deprivation protocol (see page 625, right column). Sheen teaches an “amino acid-free, glucose-free RPMI-1640” (see page 625, left column). It is well known in the art that glutamine is an amino acid. Therefore Sheen teaches a commercial medium that is free of glucose and glutamine.
To produce cell culture media deficient of single essential amino acids, Sheen teaches the amino-acid free media can be reconstituted and supplemented with individual amino acids except those to be omitted (see page 625, right column).
It would have been obvious to combine the teachings of the prior art by using a culture medium with no glutamine or sugar. Hattori teaches sugar free and low nutrient conditions. Mitcheson teaches culturing without glutamine. Sheen teaches a commercial base medium lacking glucose and glutamine. One would have been motivated to use a commercially available medium lacking sugar and amino acids, including glutamine, to achieve the sugar free and low nutrient conditions taught by Hattori. The skilled artisan would have been motivated to exclude glutamine since Mitcheson teaches glutamine free conditions avoids contamination of myocyte cultures by non-myocytes. One would have had a reasonable expectation of success since Hattori teaches commercial mediums can be used and modified to achieve the desired conditions for cardiomyocyte selection. One would have expected similar results since each reference is directed to a method of modifying medium components for cell culture. Because culturing in a medium without glutamine is rendered obvious, it would be expected to induce cell death of undifferentiated stem cells. Claim 6 is rendered obvious.
Hattori teaches cells can be selected for 24 hours under a low serum condition ([0078]). Therefore claim 10 is included in this rejection.
Hattori teaches it is effective to supplement the culture medium with a lactic acid (Lactate, 0.1-5 mM) or a pyruvic acid (0.5-5 mM) ([0071] [0072]). Therefore Hattori teaches lactate supplemented medium. Claim 30 is included in this rejection.
Hattori teaches pyruvate supplemented medium ([0072]). Therefore claim 31 is included in this rejection.
Hattori teaches human embryonic stem cells (see [0053]). Therefore claim 38 is included in this rejection.
As set forth in the rejection of claim 6 above, Hattori discloses culturing under sugar free conditions. Hattori identifies glucose as a sugar ([0064]). Claim 40 is included in this rejection.
Regarding independent claim 45: The teachings of the prior art are reiterated. The teachings of Hattori are reiterated.
While Hattori teaches culturing in a culture medium that is sugar free, the art does not teach culturing in a medium that is also free of glutamine.
The teachings of Mitcheson and Sheen are reiterated.
It would have been obvious to combine the teachings of the prior art by using a culture medium with no glutamine or sugar. Hattori teaches sugar free and low nutrient conditions. Mitcheson teaches culturing without glutamine. Sheen teaches a commercial base medium lacking glucose and glutamine. One would have been motivated to use a commercially available medium lacking sugar and amino acids, including glutamine, to achieve the sugar free and low nutrient conditions taught by Hattori. The skilled artisan would have been motivated to exclude glutamine since Mitcheson teaches glutamine free conditions avoids contamination of myocyte cultures by non-myocytes. One would have had a reasonable expectation of success since Hattori teaches commercial mediums can be used and modified to achieve the desired conditions for cardiomyocyte selection. One would have expected similar results since each reference is directed to a method of modifying medium components for cell culture. Because culturing in a medium without glutamine is rendered obvious, it would be expected to induce cell death of undifferentiated stem cells. Claim 45 is rendered obvious.
Hattori teaches selection of cardiomyocytes by culturing under sugar-free conditions ([0037]). Sugar free conditions read on the amount of sugar recited in claim 46.
Hattori teaches induced pluripotent stem cells can be used ([0059]). Therefore claim 47 is included in this rejection.
Hattori teaches embryonic stem cells (supra). Therefore claim 48 is included in this rejection.
Sheen’s medium does not contain amino acids. It is well known in the art that glutamate is an amino acid. Therefore it is not present in Sheen’s medium. Claims 49-51 are included in this rejection.
Hattori teaches lactate (supra). Claim 52 is included in this rejection.
Therefore Applicant’s invention is rendered obvious as claimed.
Claim 32 is rejected under 35 U.S.C. 103 as being unpatentable over Hattori et al. in view of Mitcheson and Sheen as applied to claim 6 above, and further in view of Whitford et al. (previously cited; Lipids in Cell Culture Media. 2008 Pages 152-154).
Claim 6 is rejected as recited in the rejection above.
While Hattori teaches the use of sugar-free culture media, the art is silent regarding the presence of fatty acids in said media. The teachings of Mitcheson and Chambers have been set forth above.
Whitford teaches supplementing cell culture systems with particular fatty acids “significantly improves performance” (page 152, middle column).
It would have been obvious to supplement the culture media disclosed by Hattori with fatty acids. One would have been motivated to do so since Whitford teaches particular fatty acids can be used in cell culture systems. One would do so to improve cell culture performance, as taught by Whitford. One would have had a reasonable expectation of success since Whitford teaches fatty acids can be added to cell culture media. One would have expected similar results since both references are directed to cell culture. Therefore claim 32 is included in this rejection as claimed.
Therefore Applicant’s Invention is rendered obvious as claimed.
37 CFR 1.132 Declaration
The Examiner acknowledges receipt of the Declaration under 37 CFR 1.132 by Dr. John Mitcheson filed on 03 September 2025. The Declarations states the Mitcheson reference contains glucose.
The Declaration under 37 CFR 1.132 is insufficient to overcome the rejection of claim 6 based upon obviousness over Hattori in view of Mitcheson and Sheen et al. as set forth in the rejection because:
The rejection is not based on the incorporation of Mitcheson’s M199 medium in Hattori.
The rejection cites why it would have been obvious to exclude glutamine in the medium taught by Hattori. Hattori teaches changing compositions of components of various commercial mediums to construct a system that selects for cardiomyocytes. Hattori explicitly excludes sugar and teaches sugar substitutes (lactic acid, pyruvic acid) “can specifically provide the cardiomyocytes with necessary nutrients” ([0072]). Hattori is silent regarding the presence of the amino acid glutamine. Mitcheson explicitly teaches excluding glutamine to select for cardiomyocytes. Sheen teaches a commercial base medium lacking glucose and glutamine. One would have been motivated to use a base medium lacking sugar and glutamine to achieve both conditions taught by Hattori. The skilled artisan would want to exclude glutamine since Mitcheson teaches glutamine free conditions avoids contamination of myocyte cultures by non-myocytes. One would have had a reasonable expectation of success since Hattori teaches commercial mediums can be used and modified to achieve the desired conditions for cardiomyocyte selection. One would have expected similar results since each reference is directed to methods of modifying mediums for cell culture.
Affidavits or declarations are provided as evidence and must set forth facts, not merely conclusions. In re Pike and Morris, 84 USPQ 235 (CCPA 1949).
Upon consideration of the facts taught by the prior art and the information submitted by the Affiant, the balance of evidence indicates that the prior art teaches the instantly claimed inventions.
APPLICANT’S ARGUMENTS
The arguments made by Applicant in the response filed on 03 September 2025 are acknowledged.
Argument 1: The Applicant argued the Declaration filed with the response is evidence the medium taught by Mitcheson contains glucose.
Response to Argument 1: The rejection is not based on incorporating Mitcheson’s medium in Hattori’s method. The rejection cites why it would have been obvious to exclude glutamine in the medium taught by Hattori. Hattori uses commercially available media to achieve the desired conditions for cardiomyocyte selection. Hattori teaches sugar free condition. Hattori teaches using a low nutrient condition. Mitcheson teaches excluding glutamine in culture mediums to select for cardiomyocytes. One would use Sheen’s commercially available medium to achieve sugar free and glutamine free conditions.
Argument 2: The Applicant argues Hattori does not teach a medium which does not contain glutamate, as recited in new claims 49-51.
Response to Argument 2: Sheen teaches a medium which does not contain any amino acids. Glutamate is an amino acid. Therefore Sheen’s medium does not contain glucose, glutamine or glutamate. The arguments are not persuasive.
Conclusion
No claims are allowed.
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) 270-8439.
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/NATALIE M MOSS/ Examiner, Art Unit 1653
/SHARMILA G LANDAU/Supervisory Patent Examiner, Art Unit 1653
