Jeffrey Mohr

US Patent:

20100197013, Aug 5, 2010

Filed:

Nov 9, 2009

Appl. No.:

12/615044

Inventors:

Timothy J. Kamp - Madison WI, US
Jianhua Zhang - Madison WI, US
Jeffrey C. Mohr - Glenview IL, US
Juan J. Depablo - Madison WI, US
Sean P. Palecek - Verona WI, US

International Classification:

C12N 5/071
C12N 5/02
C12N 5/00
C40B 40/14

US Classification:

435366, 435377, 435325, 506 20, 435374

Abstract:

A three-dimensional microwell system that supports long term pluripotent cell culture and formation of homogeneous embryoid bodies (EBs) is described. Microwell-cultured pluripotent cells remain viable and undifferentiated for several weeks in culture and maintain undifferentiated replication when passaged to Matrigel-coated, tissue culture-treated polystyrene dishes. Microwell-cultured pluripotent cells maintain pluripotency, differentiating to each of the three embryonic germ layers. Pluripotent cell aggregates released from microwells can be passaged for undifferentiated replication or differentiated to monodisperse EBs. The ability to constrain pluripotent cell growth in three dimensions advantageously provides for more efficient, reproducible culture of undifferentiated cells, high-throughput screening, and the ability to direct pluripotent cell differentiation by generating monodisperse EBs of a desired size and shape. Cardiomyocyte-rich EBs are obtained from pluripotent cells cultured in microwells of defined size and shape.