Wisconsin Alumni Research Foundation

Jianhua Zhang

Timothy Joseph Kamp

Sean Paul Palecek

Xiaojun Lance Lian

Jeffrey C Mohr

Juan J Depablo

Samira M Azarin

Methods for generating high-yield, high-purity cardiac fibroblasts are described. Differentiation methods comprising chemically defined culture conditions and methods for in vitro maintenance of human pluripotent stem cell-derived cardiac fibroblasts are also provided.

Method of differentiating human pluripotent cells into cardiac fibroblasts

Methods for culturing the pluripotent stem cells to undergo epithelial-to-mesenchymal transition and for generating high-yield, high-purity cardiomyocyte cultures from pluripotent stem cells are described. Pluripotent stem cells are cultured on a support with an overlaid matrix and, optionally, exposed to one or more factors to induce epithelial-to-mesenchymal transition and cardiogenesis.

Cardiac differentiation of human pluripotent stem cells under defined conditions using matrix overlay methods

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&#xae;-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.

Growing community of inventors

Madison, WI, United States of America

Jianhua Zhang