Patent ID: 8637309
Filing Date: 2014-01-28
Classification: C12N

Abstract:
1. A method for achieving stable, long-term culture and expansion of human multipotent stem cells in suspension culture in vitro, the method comprising: (i) attaching human multipotent stem cells to a plurality of microcarriers to form microcarrier-stem cell complexes, wherein the surface of the microcarriers is coated in a matrix comprising an extracellular matrix component; (ii) culturing the microcarrier-stem cell complexes in suspension culture; (iii) passaging the cultured cells from (ii); and (iv) repeating steps (i)-(iii) through at least 7 passages; wherein human multipotent stem cells are expanded between each passage, and wherein stem cells in the culture after step (iv) are multipotent, whereby stable, long-term culture and expansion of human multipotent stem cells is achieved in suspension in vitro, wherein the achievement of stable, long-term culture is characterized by 50% or more of the human multipotent stem cells in the culture after step (iv) having the ability to differentiate into cells of a limited number of type within a particular lineage and retaining at least one stem cell characteristic selected from the group consisting of expression of a multipotency marker, cell viability, and normal karyotype, and wherein the achievement of expansion is characterized by the number of cultured, expanded human multipotent stem cells at the end of step (ii) being at least 0.2 order of magnitude greater than the number of human multipotent stem cells attached to the microcarriers in step (i).