Defined culture medium for stem cell differentiation: applicability of serum-free conditions in the XXXX embryonic stem cell test.

@entity137 @entity19 @entity620 @entity8385 @entity7739
The embryonic stem cell test (EST) is a validated method to assess the developmental @entity137 potency of chemicals. It was developed to reduce animal use and allow faster testing for hazard assessment. The cells used in this method are maintained and differentiated in media containing foetal calf serum. This animal product is of considerable variation in quality, and individual batches require extensive testing for their applicability in the EST. Moreover, its production involves a large number of foetuses and possible animal suffering. We demonstrate the serum-free medium and feeder cell-free maintenance of the @entity19 embryonic stem cell line D3 and investigate the use of specific growth factors for induction of cardiac differentiation. Using a combination of @entity8385 , @entity7739 , activin A and @entity620 , embryoid bodies efficiently differentiated into contracting myocardium. Additionally, examining levels of intracellular marker proteins by flow cytometry not only confirmed differentiation into cardiomyocytes, but demonstrated significant differentiation into neuronal cells in the same time frame. Thus, this approach might allow for simultaneous detection of developmental effects on both early mesodermal and neuroectodermal differentiation. The serum-free conditions for maintenance and differentiation of D3 cells described here enhance the transferability and standardisation and hence the performance of the EST.
