Composition and method for enabling proliferation of pluripotent stem cells
The present disclosure is directed to the development of compositions, such as extracellular matrices, and processes for using the same, for culturing stem cells in vitro in an undifferentiated state. In this regard, it has been discovered that when pluripotent mouse and human embryonic stem cells are cultured on plates coated with recombinant laminin-10 (laminin-511) or laminin-5 (laminin-322), or their functional domains, the embryonic stem cells proliferated and maintained their pluripotency.
1. A method for enabling proliferation of pluripotent murine embryonic stem cells grown in vitro, comprising: providing an extracellular matrix comprising (i) a laminin selected from the group consisting of recombinant laminin 332 (laminin-5) and recombinant laminin 511 (laminin-10), and (ii) a medium that comprises beta fibroblast growth factor (bFGF); and coating the extracellular matrix with pluripotent murine embryonic stem cells.
2. The method of claim 1 , wherein the extracellular matrix is devoid of any differentiation inhibitor.
3. A composition for enabling proliferation of pluripotent stem cells in-vitro for greater than 14 days consisting of a layer of (i) a laminin selected from the group consisting of recombinant laminin 332 (laminin-5) and recombinant laminin 511 (laminin-10) and (ii) beta fibroblast growth factor (bFGF).