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Controlled, Scalable Embryonic Stem Cell Differentiation Culture

^a Institute of Biomaterials and Biomedical Engineering, Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Canada;
^b Biotechnology Interdisciplinary Unit, Technion-Israel Institute of Technology, Haifa, Israel;
^c Department of Obstetrics and Gynecology, Rambam Medical Center, Haifa, Israel;
^d Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel

Key Words. Stem cell culture • Encapsulation • Embryonic stem cells • Embryoid body • Scalable

Peter W. Zandstra, Ph.D., Institute of Biomaterials and Biomedical Engineering, Room 407, Roseburgh Building, 4 Taddle Creek Road, Toronto, Ontario, M5S 3G9, Canada. Telephone: 416-978-8888; Fax: 416-978-4317; e-mail: peter.zandstra{at}utoronto.ca

Embryonic stem (ES) cells are of significant interest as a renewable source of therapeutically useful cells. ES cell aggregation is important for both human and mouse embryoid body (EB) formation and the subsequent generation of ES cell derivatives. Aggregation between EBs (agglomeration), however, inhibits cell growth and differentiation in stirred or high-cell-density static cultures. We demonstrate that the agglomeration of two EBs is initiated by E-cadherin-mediated cell attachment and followed by active cell migration. We report the development of a technology capable of controlling cell-cell interactions in scalable culture by the mass encapsulation of ES cells in size-specified agarose capsules. When placed in stirred-suspension bioreactors, encapsulated ES cells can be used to produce scalable quantities of hematopoietic progenitor cells in a controlled environment.

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