Protocol abstract

Nature Protocols 3, 768 - 776 (2008)
Published online: 10 April 2008 | doi:10.1038/nprot.2008.42

Subject Categories: Cell and developmental biology | Cell and tissue culture

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

Elizabeth S Ng1, Richard Davis1, Edouard G Stanley1 & Andrew G Elefanty1

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.

  1. Monash Immunology and Stem Cell Laboratories, STRIP 1, Building 75, Level 3, Monash University, Clayton, Victoria 3800, Australia.

Correspondence to: Andrew G Elefanty1 e-mail: