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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 21 November 2025 has been entered.
DETAILED OFFICE ACTION
This Office Action is in response to the papers filed on 21 November 2025.
CLAIMS UNDER EXAMINATION
Claims 1 and 4-6 have been examined on their merits.
PRIORITY
KR10-2019-0098344 filed on 12 August 2019 is acknowledged. A certified translation has not been provided.
WITHDRAWN OBJECTIONS
The objection to claim 14 has been withdrawn.
WITHDRAWN REJECTIONS
The previous rejections have been withdrawn due to claim amendment.
REJECTIONS
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 1 and 4-6 are rejected under 35 U.S.C. 103 as being unpatentable over Lee et al. (previously cited; Composition and method for differentiation adult stem cells into chondrocytes KR20170108325A. 2017-09-27) in view of Muttigi et al. (previously cited; Matrilin-3 codelivery with adipose-derived mesenchymal stem cells promotes articular cartilage regeneration in a rat osteochondral defect model. J. Tissue Eng Regen Med. 2018 Mar;12(3):667-675; first published 29 May 2017) and Yamaguchi et al. (previously cited; Mesenchymal stem cell spheroids exhibit enhanced in-vitro and in-vivo osteoregenerative potential. BMC Biotechnology (2014) 14:105).
Lee teaches a composition and method for differentiating adult stem cells into chondrocytes using matrilin-3 (Technical Field section, first two sentences). Adult stem cells are cultured in the presence of MATN3 protein (see page 2, 7th section of page). Cells can be cultured for 3 days (72 hours) (i.e., about 80 hours) (last paragraph on page 2). The art teaches “a three-dimensional culture method” can be applied (first paragraph of page 3). The art teaches three-dimensional culture refers to a method of culturing a cell such that a monolayer is grown to form a three-dimensional solid form, such as a “sphere” (same cited section). Lu treats cells with 20 ng/ml MATN3 (Figure 4).
Lee does not teach 10 ng/ml MATN3.
Lee is silent regarding the number of cells used to perform the culture.
Muttigi et al. teach a method of promoting articular cartilage regeneration in a rat
(Abstract). Mesenchymal stem cells from adipose are isolated (see
section 2.1). Isolated cells are suspended in culture medium (see page 668, right
column). Muttigi treats cells with 10 ng/ml, 20 ng/ml or 50 ng/ml matrilin-3 (see page 668, right column, section 2.2 and 2.3; see page 669, left column, section 2.4; see Figure 3). Muttigi teaches these concentrations do not induce cytotoxicity (see section 3.2). Muttigi teaches 10 ng/ml matrilin-3 increased concentration of chondrogenic markers relative to untreated cells (see Group 2 in Figure 2). Figure 3 discloses treatment with 10 ng/ml and 20 ng/ml matrilin-3 promote chondrogenic differentiation of Ad-MSCs in vitro using pellet culture (see text of Figure 3). As evidenced by the specification, pellet culture is 3D cell culture.
Muttigi teaches the effect of matrilin-3 is dependent on its concentration, with higher concentrations having a reduced regenerative potential when matrilin-3 is co-delivered with adipose derived mesenchymal stem cells (see page 674, right column, lines 1-10-15).
Yamaguchi investigates the effects of seeding density on mesenchymal stem cell (MSC) spheroid formation (see page 2, left column, second paragraph of Results section). 100 cells are seeded per well (same cited section). The art teaches spheroid size can be controlled by cell density (see page 2, left column, second paragraph of Results section). Yamaguchi uses 96 well plates (hence, microwells; page 8, right column, first paragraph).
The art also teaches the following:
Yamaguchi teaches mesenchymal stem cells (MSCs) are a favored cell source for regenerative medicine because of their multilineage potential (Abstract).
MSCs are capable of self-renewal and differentiation into several mesenchymal lineages in-vitro and in-vivo, including bone, fat, cartilage, and skeletal muscles (see first paragraph of Background on page 1). MSCs are commonly cultured as a two-dimensional (2D) monolayer using conventional tissue-culture techniques. These 2D-monolayer techniques inadequately reproduce the in-vivo microenvironment of stem cells, established by extrinsic and intrinsic cellular signals and have a profound influence on their biological functions (see page 1, right column, first paragraph).
Culture of MSCs in multicellular spheroids, more accurately mimics the in-vivo microenvironment; thus, resolving this problem (Abstract; see page 1, last paragraph).
It would have been obvious to try using 10 ng/ml matrilin-3 in the method taught by Lee. One would have been motivated to do so since Muttigi teaches 10 ng/ml induces chondrogenic differentiation of adipose-tissue-derived mesenchymal stem cells. Because Muttigi teaches Ad-MSCs treated with higher concentrations of matrilin-3 have a reduced regenerative effect, the skilled artisan would use the lowest dose that is able to produce the desired chondrogenic effect. One would have had a reasonable expectation of success since Muttigi teaches Ad-MSCs can be successfully treated with 10 ng/ml matrilin-3. One would have expected similar results since both references treat the same cells with matrilin-3 to promote chondrogenesis.
It would have been obvious to culture 100 stem cells to perform 3D culture. Lee teaches 3D culture of mesenchymal stem cells and Yamaguchi uses 100 mesenchymal stem cells per microwell to perform 3D culture. One would have been motivated to use 100 cells per well to produce smaller spheroids, as taught by Yamaguchi. One would have had a reasonable expectation of success since Lee teaches formation of a sphere, and Yamaguchi forms a spheroid (a sphere) using 100 mesenchymal stem cells. One would have expected similar results since both references culture mesenchymal stem cells using 3D culture. Therefore claim 1 is rendered obvious.
Lee teaches human adult stem cells (see page 2, third paragraph). Therefore claim 4 is rendered obvious.
Lee teaches adipose derived stem cells (see page 2, second paragraph). Therefore claim 5 is rendered obvious.
Lee teaches the three-dimensional culture is preferably a pellet culture (first paragraph of page 3). Therefore claim 6 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 21 November 2025 are acknowledged. The Applicant argues the prior art does not teach the limitations recited in amended claim 1. The Applicant argues 10 ng/ml matrilin-3 produces an unexpected result. The arguments state it was demonstrated 10 ng/ml matrilin-3 (Group 2) resulted in the greatest gene content and the highest synthesis degree (Figure 5B) and achieved maximum expressions of cartilage differentiation markers (collagen 2 and aggrecan (Figure 5C). The Applicant argues no treatment (Group 1) and Groups 3 and 4 exhibit reduced expression of differentiation markers.
The Applicant argues Yamaguchi is directed to osteogenesis and does not teach matrilin-3.
EXAMINER’S RESPONSE
New grounds of rejection have been made to address amended claim 1.
Claim 1 recites culturing in a range of about 80 hours to about 130 hours.
The experiments in Figure 5C demonstrate increased COL2A expression and ACAN expression at day 5 (120 hours). At Day 3 (72 hours, i.e., about 80 hours), Group 2 (10 ng/ml) results in decreased expression of collagen 2 and aggrecan while 20 ng/ml demonstrates increased expression. At [0070], the PG Pub states “when the human adipose-derived stem cells were isolated, the MATN-3 concentration was 10 ng/ml, and the MATN-3 priming period was 5 days, the greatest gene content and the highest synthesis degree were resulted (see FIG. 5B), and that the cartilage differentiation markers (e.g., collagen 2 and aggrecan) were expressed to the maximum”.
The experiments pointed to by the Applicant do not demonstrate an unexpected result for the range of time recited in claim 1.
As set forth above, treating Ad-MSCs with 10 ng/ml matrilin-3 to promote chondrogenesis was known in the prior art. Therefore the claimed concentration would be expected to produce the effects recited by the Applicant.
The arguments directed to Yamaguchi are not persuasive. Yamaguchi is not relied upon to teach matrilin-3. Lee and Muttigi teach matrilin-3. Yamaguchi is not relied upon to teach chondrogenic differentiation. As set forth above, Yamaguchi teaches MSCs are capable of self-renewal and differentiation into several mesenchymal lineages in-vitro and in-vivo, including cartilage (hence, chondrogenesis). Yamaguchi teaches culture of MSCs in multicellular spheroids more accurately mimics the in-vivo microenvironment.
Yamaguchi is relied upon because it teaches Ad-MSCs have the ability to form spheroids with 100 cells in microwells.
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