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

Biotechnol Bioeng. 2012 Apr;109(4):853-66. doi: 10.1002/bit.24375. Epub 2011 Dec 2.


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.

Publication types

  • Editorial
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Differentiation / analysis
  • Batch Cell Culture Techniques*
  • Cell Aggregation
  • Cell Cycle
  • Cell Differentiation
  • Cell Division
  • Cell Line / cytology
  • Cell Line / drug effects
  • Coculture Techniques
  • Culture Media
  • Endoderm / cytology*
  • Fibroblasts / metabolism
  • Hepatocytes / cytology
  • Humans
  • Mice
  • Pancreas / cytology
  • Pluripotent Stem Cells / cytology*
  • Suspensions
  • Tissue Engineering / methods*


  • Antigens, Differentiation
  • Culture Media
  • Suspensions