MOUSE WHOLE-EMBRYO CULTURE IN SERUM DILUTED WITH WAYMOUTH MEDIUM - A STUDY OF VALPROIC ACID TERATOGENICITY

The culture of whole post-implantation rodent embryos has become an important tool in developmental biology and toxicology. Since the establishment of this system, rat serum has been the main culture medium used. In this study, we demonstrate that medium composed of 50% serum and 50% chemically defined medium (Waymouth 705/1) permits satisfactory development of mouse embryos over a 26-hour period. In addition, our data demonstrate that, with less than 50% serum, the frequency of malformations increases and growth and differentiation decrease with dilution in a dose-dependent way. Little information is available in the literature on the types of abnormalities induced by inadequate dilutions of serum. This study shows that a chemically defined medium, supplemented with amounts of serum that are below threshold levels, interferes mainly with the normal development of the head (neural tube, eyes and maxillary processes) and with the growth of the embryo. The second part of this work compares the teratogenicity of valproic acid (VPA) in this new medium to that in undiluted serum. We show that the effects of VPA are similar in terms of the type of alterations observed, the reduction of growth and the differentiation. However, the concentrations necessary to induce these effects in Waymouth/human serum/rat serum are half of those needed in human serum/rat serum.