US 12,441,984 B2
Isolation, preservation, and expansion of canine umbilical cord mesenchymal stromal cells
Mark L. Weiss, Manhattan, KS (US); and Adrienne Wright, Manhattan, KS (US)
Assigned to Kansas State University Research Foundation, Manhattan, KS (US)
Appl. No. 17/774,968
Filed by Kansas State University Research Foundation, Manhattan, KS (US)
PCT Filed Nov. 5, 2020, PCT No. PCT/US2020/059143
§ 371(c)(1), (2) Date May 6, 2022,
PCT Pub. No. WO2021/092199, PCT Pub. Date May 14, 2021.
Claims priority of provisional application 62/932,844, filed on Nov. 8, 2019.
Prior Publication US 2022/0380731 A1, Dec. 1, 2022
Int. Cl. A61K 35/12 (2015.01); C12N 5/0775 (2010.01)
CPC C12N 5/0668 (2013.01) [C12N 2500/32 (2013.01); C12N 2501/115 (2013.01); C12N 2509/00 (2013.01); C12N 2513/00 (2013.01); C12N 2533/50 (2013.01); C12N 2533/70 (2013.01)] 16 Claims
OG exemplary drawing
 
1. A method of expanding isolated canine umbilical cord mesenchymal stromal cells (UC-MSCs) from umbilical cord tissue, said method comprising:
adding said isolated canine UC-MSCs from umbilical cord tissue and cell culture media to a first adherent cell culture substrate, said cell culture media being supplemented with FBS, glutamine, and fibroblast growth factor-basic, wherein said first adherent cell culture substrate has a cell culture surface that is coated with a plurality of cell adhesion molecules selected from the group consisting of gelatin, laminin, fibronectin, poly-lysine, vibronectin, collagen, and/or fibrin;
incubating said isolated canine UC-MSCs under conditions to yield cultured canine UC-MSCs;
transferring said cultured canine UC-MSCs to a second adherent cell culture substrate, wherein said transferring comprising subjecting said cultured canine UC-MSCs to a dissociation reagent to detach said cells from said first adherent cell culture substrate and moving said detached cells to said second adherent cell culture substrate, wherein said second adherent cell culture substrate has a cell culture surface that is coated with a plurality of cell adhesion molecules selected from the group consisting of gelatin, laminin, fibronectin, poly-lysine, vibronectin, collagen, and/or fibrin; and
incubating said isolated canine UC-MSCs under conditions to yield expanded canine UC-MSCs, wherein said expanded canine UC-MSCs have a cell population that is greater than said isolated canine UC-MSCs.