In vitro culture of peri-gastrulation embryos of a macropodid marsupial.

Peri-gastrulation stage tammar wallaby embryos were cultured for up to 78 h in either Dulbecco's Modified Eagle's Medium or Medium 199, in air/6% CO2 or 95% O-2/5% CO2, and with added fetal calf or wallaby serum. There was little difference between the two media or sera sources, but development was markedly superior for embryos cultured in 95% O-2/5% CO2. Many embryos survived even prolonged culture periods up to and over 70 h, and although development continued throughout the culture period, the embryos as a whole became increasingly abnormal. Embryos explanted at the primitive streak/ regressing node stages performed better in vitro than embryos explanted at earlier or later stages. The embryo that developed the furthest had a newly formed node at the initiation of culture and after 64 h in vitro it had developed forelimb ridges, fused, beating heart tubes and mesonephric ducts. Thus high oxygen appears to be the critical component of the culture system for optimal development of primitive streak stage tammar embryos. These results provide a basis for developing culture conditions for longer term development of marsupial embryos in vitro.