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Attachment and growth of human embryonic stem cells on microcarriers

Journal Article


Abstract


  • The use of human embryonic stem cells (hESCs) for cell-based therapies will require large quantities of genetically stable pluripotent cells and their differentiated progeny. Traditional hESC propagation entails adherent culture and is sensitive to enzymatic dissociation. These constraints hamper modifying method from 2-dimensional flat-bed culture, which is expensive and impractical for bulk cell production. Large-scale culture for clinical use will require innovations such as suspension culture for bioprocessing. Here we describe the attachment and growth kinetics of both murine embryonic stem cells (mESCs) and hESCs on trimethyl ammonium-coated polystyrene microcarriers for feeder-free, 3-dimensional suspension culture. mESCs adhered and expanded according to standard growth kinetics. For hESC studies, we tested aggregate (collagenase-dissociated) and single-cell (TrypLE™-dissociated) culture. Cells attached rapidly to beads followed by proliferation. Single-cell cultures expanded 3-fold over approximately 5 days, slightly exceeding that of hESC aggregates. Importantly, single-cell cultures were maintained through 6 passages with a 14-fold increase in cell number while still expressing the undifferentiated markers Oct-4 and Tra 1-81. Finally, hESCs retained their capacity to differentiate towards pancreatic, neuronal, and cardiomyocyte lineages. Our studies provide proof-of-principle of suspension-based expansion of hESCs on microcarriers, as a novel, economical and practical feeder-free means of bulk hESC production.

Authors


  •   Phillips, Blaine W. (external author)
  •   Horne, Rachel (external author)
  •   Lay, Tan S. (external author)
  •   Rust, William L. (external author)
  •   Teck, Tan Thong (external author)
  •   Crook, Jeremy M.

Publication Date


  • 2008

Citation


  • Phillips, B. Wesley., Horne, R., Lay, T. Suat., Rust, W. Lanthrop., Teck, T. Thong. & Crook, J. Micah. (2008). Attachment and growth of human embryonic stem cells on microcarriers. Journal of Biotechnology, 138 (1-2), 24-32.

Scopus Eid


  • 2-s2.0-53549120197

Ro Metadata Url


  • http://ro.uow.edu.au/aiimpapers/1188

Has Global Citation Frequency


Number Of Pages


  • 8

Start Page


  • 24

End Page


  • 32

Volume


  • 138

Issue


  • 1-2

Place Of Publication


  • Netherlands

Abstract


  • The use of human embryonic stem cells (hESCs) for cell-based therapies will require large quantities of genetically stable pluripotent cells and their differentiated progeny. Traditional hESC propagation entails adherent culture and is sensitive to enzymatic dissociation. These constraints hamper modifying method from 2-dimensional flat-bed culture, which is expensive and impractical for bulk cell production. Large-scale culture for clinical use will require innovations such as suspension culture for bioprocessing. Here we describe the attachment and growth kinetics of both murine embryonic stem cells (mESCs) and hESCs on trimethyl ammonium-coated polystyrene microcarriers for feeder-free, 3-dimensional suspension culture. mESCs adhered and expanded according to standard growth kinetics. For hESC studies, we tested aggregate (collagenase-dissociated) and single-cell (TrypLE™-dissociated) culture. Cells attached rapidly to beads followed by proliferation. Single-cell cultures expanded 3-fold over approximately 5 days, slightly exceeding that of hESC aggregates. Importantly, single-cell cultures were maintained through 6 passages with a 14-fold increase in cell number while still expressing the undifferentiated markers Oct-4 and Tra 1-81. Finally, hESCs retained their capacity to differentiate towards pancreatic, neuronal, and cardiomyocyte lineages. Our studies provide proof-of-principle of suspension-based expansion of hESCs on microcarriers, as a novel, economical and practical feeder-free means of bulk hESC production.

Authors


  •   Phillips, Blaine W. (external author)
  •   Horne, Rachel (external author)
  •   Lay, Tan S. (external author)
  •   Rust, William L. (external author)
  •   Teck, Tan Thong (external author)
  •   Crook, Jeremy M.

Publication Date


  • 2008

Citation


  • Phillips, B. Wesley., Horne, R., Lay, T. Suat., Rust, W. Lanthrop., Teck, T. Thong. & Crook, J. Micah. (2008). Attachment and growth of human embryonic stem cells on microcarriers. Journal of Biotechnology, 138 (1-2), 24-32.

Scopus Eid


  • 2-s2.0-53549120197

Ro Metadata Url


  • http://ro.uow.edu.au/aiimpapers/1188

Has Global Citation Frequency


Number Of Pages


  • 8

Start Page


  • 24

End Page


  • 32

Volume


  • 138

Issue


  • 1-2

Place Of Publication


  • Netherlands