Canine iPS cells and method of producing same
AssigneeKyoto University
InventorsTatsuo Nakamura, Hidenori Shimada
Life of this patent
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Abstract
Provided are a method of producing canine iPS cells, comprising (a) the step of bringing into contact with each other a canine somatic cell and a nuclear reprogramming factor, and (b) the step of culturing the cell in a medium containing at least one substance selected from the group consisting of a mitogen-activated protein kinase kinase inhibitor, an activin receptor-like kinase inhibitor, a glycogen synthase kinase inhibitor, a L-type calcium channel agonist and a DNA methylation inhibitor, and a leukemia inhibitory factor, and canine iPS cells that can be obtained by the method.
Claims7
A method of altering the differentiation state of a canine somatic cell to a less differentiated state, comprising:
- (a) transfecting the canine somatic cell with a retroviral vector comprising nucleic acids encoding each of Oct3/4, Sox2, Klf4, and c-Myc operably linked to a promoter, and
- (b) culturing the cell in a medium containing a mitogen-activated protein kinase kinase inhibitor, an activin receptor-like kinase inhibitor, a glycogen synthase kinase inhibitor, a histone deacetylase inhibitor, a basic fibroblast growth factor, and a leukemia inhibitory factor to provide a resultant canine cell,
- wherein the resultant canine cell (i) has an altered fate potential relative to the starting canine somatic cell, (ii) has a morphology similar to a canine ES cell, (iii) expresses Oct3/4, Sox2, Sall4, SSEA-4, TRA-1-60, and TRA-1-81 genes, (iv) is positive for alkaline phosphatase activity, (v) maintains normal karyotype, and (vi) has a potential to differentiate into a βIII-tubulin-expressing ectodermal cell, a FLK1-expressing mesodermal cell, and an α-fetoprotein-expressing endodermal cell.
Dependencies are inferred from the claim text.