Patent ID: 8716018
Filing Date: 2014-05-06
Classification: C12N

Abstract:
1. A method for achieving stable, long-term culture and expansion of human embryonic stem cells, or human induced pluripotent stem cells, in suspension in vitro, the method comprising: (i) attaching human embryonic stem cells or human induced pluripotent 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 stem cells in the culture after step (iv) are pluripotent, thus achieving stable, long-term culture and expansion of human embryonic stem cells or human induced pluripotent stem cells in suspension in vitro, wherein the achievement of stable, long-term culture is characterized by 50% or more of the stem cells in the culture after step (iv) having the ability to differentiate into endoderm, ectoderm, and mesoderm and retaining at least one stem cell characteristic selected from the group consisting of expression of a pluripotency marker, cell viability, and normal karyotype, and wherein the achievement of expansion is characterized by the number of cultured, expanded cells at the end of step (ii) being at least 0.2 order of magnitude greater than the number of cells attached to the microcarriers in step (i).