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Bioprocess and Biosystems Engineering

, Volume 27, Issue 6, pp 381–387 | Cite as

Perfusion cultures of human embryonic stem cells

  • Wey Jia Fong
  • Heng Liang Tan
  • Andre Choo
  • Steve Kah Weng OhEmail author
Original papers

Abstract

Human embryonic stem cells (hESC) are self-renewing pluripotent cells capable of differentiating into cells representative of all three embryonic germ layers. Hence, they hold great potential for regenerative medicine. However, significant cell numbers are required to fulfill their potential therapeutic applications. In this study, perfusion with supplemented conditioned media (SCM), produced by mouse embryonic fibroblasts (MEF), was adopted to improve cell densities of hESC cultures. Perfusion enhanced hESC numbers by 70% compared to static conditions, on both organ culture dish (OCD) and petridish cultures. All cultures maintained healthy expression of the pluripotent marker, Oct-4 transcription factor. In vivo, perfused hESC formed teratomas in severe combined immunodeficiency (SCID) mice models that represent the three embryonic germ layers. When SCM was produced with lower concentrations of MEF, hESC densities and Oct-4 levels were reduced. Hence, perfusion with SCM is a potential feeding method for scale-up production of hESC.

Keywords

Perfusion Human embryonic stem cell Conditioned media Pluripotency Oct-4 

Abbreviations

hESC

Human embryonic stem cell

SCID

Severe combined immunodeficiency

CM

Conditioned media

SCM

Supplemented conditioned media

MEF

Mouse embryonic fibroblast

PET

Polyethylene terepthalate

mAb

Monoclonal antibody

mESC

Mouse embryonic stem cell

FACS

Fluorescence activated cell sorter

Notes

Acknowledgements

This work was generously supported by the Agency for Science Technology and Research (A*STAR), Singapore. We thank Angela Chin and Jayanthi Padmanabhan very much for their help with teratoma sectioning and FACS. We thank Dr. Victor Wong for critical review of this manuscript.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Wey Jia Fong
    • 1
  • Heng Liang Tan
    • 1
  • Andre Choo
    • 1
  • Steve Kah Weng Oh
    • 1
    Email author
  1. 1.Bioprocessing Technology InstituteCentrosSingapore

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