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

被引：228

作者：

Ng, Elizabeth S. ^{[1
]}

Davis, Richard ^{[1
]}

Stanley, Edouard G. ^{[1
]}

Elefanty, Andrew G. ^{[1
]}

机构：

[1] Monash Univ, Monash Immunol & Stem Cell Labs, Clayton, Vic 3800, Australia

来源：

NATURE PROTOCOLS | 2008年 / 3卷 / 05期

基金：

英国医学研究理事会;

关键词：

D O I：

10.1038/nprot.2008.42

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：768 / 776

页数：9