In vitro production of expanded potential stem cells
AssigneeGenome Research Limited
InventorsPentao LIU, David RYAN, Xuefei GAO, Wei WANG, Jian YANG
Life of this patent
Estimating the term…
Abstract
A culture medium is provided which is capable of establishing expanded potential stem cell (EPSC) lines which resemble naive or ground state ES cells, but are also able to differentiate into placenta trophoblasts and the embryo proper. Methods are provided using the medium for the in vitro conversion and maintenance of cells, including pluripotent cells into EPSCs.
Claims10
1-49. (canceled)
A method for producing a population of expanded potential stem cells (EPSCs) which comprises:
- (i) Providing a population of pluripotent cells, wherein the pluripotent cells are obtained by
- (a) Providing a population of non-pluripotent cells
- (b) Reprogramming the population of non-pluripotent cells by introducing pluripotency genes or proteins into the cells thereby producing a population of pluripotent stem cells;
- and
- (ii) Culturing the population in an expanded potential stem cell medium (EPSCM), wherein the EPSCM comprises one or more of a Src Kinase Family (SKF) inhibitor and a tankyrase inhibitor, thereby producing a population of EPSCs, wherein the pluripotency genes or proteins are Oct4, Sox2, Klf4, cMyc, Lrh1 and Rarg, and optionally Nanog and Lin28.
An expanded potential stem cell culture medium (EPSCM) comprising a basal cell nutrient medium and inhibitors comprising two or more of a Src Kinase family (SFK) inhibitor, a GSK3 inhibitor and a Wnt inhibitor.
A method for sorting a population of nave and primed pluripotent stem cells (PSCs) into a homogeneous population of expanded potential stem cells (EPSCs), the method comprising culturing the PSCs in an expanded potential stem cell medium (EPSCM), wherein the EP SCM comprises at least two of a Src Kinase family (SFK) inhibitor a GSK3 inhibitor, and a Wnt inhibitor, thereby producing a homogeneous population of EPSCs.
A method of making a transgenic non-human animal comprising:
- (i) generating a population of expanded potential stem cells (EPSCs),
- (ii) injecting the EPSCs generated in (i) into a blastocyst to generate a chimera, and
- (iii) culturing the blastocyst under conditions suitable to develop into a non-human animal.
Dependencies are inferred from the claim text.