Editors' Choice

Rational bioprocess design for human pluripotent stem cell expansion and endoderm differentiation based on cellular dynamics

  1. Mark D. Ungrin1,2,3,
  2. Geoff Clarke1,2,
  3. Ting Yin1,2,
  4. Sylvia Niebrugge1,2,
  5. M. Cristina Nostro3,
  6. Farida Sarangi3,
  7. Geoffrey Wood4,
  8. Gordon Keller3,
  9. Peter W. Zandstra1,2,3,5,6,*

Article first published online: 2 DEC 2011

DOI: 10.1002/bit.24375

Issue

Biotechnology and Bioengineering

Biotechnology and Bioengineering

Volume 109, Issue 4, pages 853–866, April 2012

Additional Information

How to Cite

Ungrin, M. D., Clarke, G., Yin, T., Niebrugge, S., Nostro, M. C., Sarangi, F., Wood, G., Keller, G. and Zandstra, P. W. (2012), Rational bioprocess design for human pluripotent stem cell expansion and endoderm differentiation based on cellular dynamics. Biotechnol. Bioeng., 109: 853–866. doi: 10.1002/bit.24375

Author Information

  1. 1

    Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada

  2. 2

    The Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Rm 1116, Toronto, Ontario, Canada M5S 3E1 416-978-8888; telephone: 416-978-8888; fax: 416-978-4317

  3. 3

    McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada

  4. 4

    Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada

  5. 5

    Heart & Stroke Richard Lewar Centre of Excellence, University of Toronto, Toronto, Ontario, Canada

  6. 6

    Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada

*Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.

Publication History

  1. Issue published online: 21 FEB 2012
  2. Article first published online: 2 DEC 2011
  3. Manuscript Accepted: 7 NOV 2011
  4. Manuscript Revised: 1 NOV 2011
  5. Manuscript Received: 2 SEP 2011

Funded by

  • NIH. Grant Number: UO1 DK089561
  • Juvenile Diabetes Research Foundation. Grant Number: 41-2009-765
  • Canadian Institutes of Health Research. Grant Number: MOP-57885
  • McLaughlin Centre for Molecular Medicine

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Keywords:

  • bioprocess;
  • pluripotent stem cells;
  • differentiation;
  • endoderm;
  • expansion

Abstract

We present a predictive bioprocess design strategy employing cell- and molecular-level analysis of rate-limiting steps in human pluripotent stem cell (hPSC) expansion and differentiation, and apply it to produce definitive endoderm (DE) progenitors using a scalable directed-differentiation technology. We define a bioprocess optimization parameter (L; targeted cell Loss) and, with quantitative cell division tracking and fate monitoring, identify and overcome key suspension bioprocess bottlenecks. Adapting process operating conditions to pivotal parameters (single cell survival and growth rate) in a cell-line-specific manner enabled adherent-equivalent expansion of hPSCs in feeder- and matrix-free defined-medium suspension culture. Predominantly instructive differentiation mechanisms were found to underlie a subsequent 18-fold expansion, during directed differentiation, to high-purity DE competent for further commitment along pancreatic and hepatic lineages. This study demonstrates that iPSC expansion and differentiation conditions can be prospectively specified to guide the enhanced production of target cells in a scale-free directed differentiation system. Biotechnol. Bioeng. 2012; 109:853–866. © 2011 Wiley Periodicals, Inc.

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