In vitro development of the mammalian embryo



Normal growth and differentiation of mammalian embryos in vitro during the preimplantation period appear to be dependent upon the availability of appropriate metabolic substrates. For preimplantation embryos, defined conditions of culture have been achieved only in a few laboratory species. There is now evidence that differentiation factors isolated from fetal calf serum and human placental cord serum may promote further development of blastocysts. Postimplantation rat and mouse embryos can be cultured during the organogenesis period with rat or human sera in roller bottles. The embryonic differentiation of the rat at this stage of development is progressively retarded in such cultures with male rat serum. The embryonic development is not improved, even in sera obtained from rats at different days of gestation (12, 15–16, and 20–21). Inability to grow placental tissues simultaneously with embryos, accumulation of unfavorable substances, and rapid depletion of nutrients contribute to the retardation of embryonic growth. To improve growth and differentiation of conceptuses, a continuous culture system with the possibility of infusion of increasing concentrations of oxygen in the roller bottle gas atmosphere has been developed. This improved method allows considerable continuous growth and differentiation from the neurula stage with development of numerous primary organs.

Utilizing these in vitro culture methods during pre- and postimplantation periods, it is now possible to assess embryotoxic or teratogenic potential of drugs and chemical agents. The postimplantation culture procedure allows a more precise assessment of mechanisms associated with anomalous embryonic differentiation. Bioactivation of teratogens and effects of active toxic metabolites on organ primordium differentiation have been shown by combining embryo culture with a hepatic microsomal activating system. Microinjection of teratogens and cells into conceptus compartments is being used to elucidate specific anomalous differentiation processes.