A protocol describing the use of a recombinant protein-based, animal product-free medium (APEL) for human embryonic stem cell differentiation as spin embryoid bodies (original) (raw)

Nature Protocols volume 3, pages 768–776 (2008)Cite this article

Abstract

In order to promote the uniform and reproducible differentiation of human embryonic stem cells (HESCs) in response to exogenously added growth factors, we have developed a method (spin embryoid bodies (EBs)) that uses a recombinant protein-based, animal product-free medium in which HESCs are aggregated by centrifugation to form EBs. In this protocol we describe the formulation of this medium, denoted APEL (Albumin Polyvinylalcohol Essential Lipids), and its use in spin EB differentiation of HESCs. We also describe a more economical variant, BPEL (Bovine Serum Albumin (BSA) Polyvinylalchohol Essential Lipids), in which BSA replaces the recombinant human albumin. The integration of a medium that includes only defined and recombinant components with a defined number of cells to initiate EB formation results in a generally applicable, robust platform for growth factor-directed HESC differentiation.

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Acknowledgements

This work was supported by the Australian Stem Cell Centre and the National Health and Medical Research Council of Australia (NHMRC). A.G.E. is a Senior Research Fellow of the NHMRC.

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Authors and Affiliations

  1. Monash Immunology and Stem Cell Laboratories, STRIP 1, Building 75, Level 3, Monash University, Clayton, 3800, Victoria, Australia
    Elizabeth S Ng, Richard Davis, Edouard G Stanley & Andrew G Elefanty

Authors

  1. Elizabeth S Ng
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  2. Richard Davis
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  3. Edouard G Stanley
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  4. Andrew G Elefanty
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Corresponding author

Correspondence toAndrew G Elefanty.

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Ng, E., Davis, R., Stanley, E. et al. A protocol describing the use of a recombinant protein-based, animal product-free medium (APEL) for human embryonic stem cell differentiation as spin embryoid bodies.Nat Protoc 3, 768–776 (2008). https://doi.org/10.1038/nprot.2008.42

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