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Perfusion cultures of human embryonic stem cells

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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.

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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

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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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Authors and Affiliations

  1. Bioprocessing Technology Institute, 20, Biopolis Way, Centros, #06-01, Singapore, 138668

    Wey Jia Fong, Heng Liang Tan, Andre Choo & Steve Kah Weng Oh

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  1. Wey Jia Fong
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  2. Heng Liang Tan
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  3. Andre Choo
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  4. Steve Kah Weng Oh
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Correspondence to Steve Kah Weng Oh.

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Fong, W.J., Tan, H.L., Choo, A. et al. Perfusion cultures of human embryonic stem cells. Bioprocess Biosyst Eng 27, 381–387 (2005). https://doi.org/10.1007/s00449-005-0421-5

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  • DOI: https://doi.org/10.1007/s00449-005-0421-5

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