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In vitro Culture of Mammalian Embryos

University of Pennsylvania, Philadelphia^,2

Abstract

Although some information is available on the nutrient requirements of preimplantation stages of mammalian embryos, much more work is required before we have a comprehensive knowledge of what these embryos need to develop in vitro, either for short periods of time during transfer or extended periods of time during manipulation. It seems logical to make the largest number and most detailed studies on the embryos of laboratory animals, and then to attempt to extrapolate this information to the embryos of the larger species. Certainly, if a good foundation of knowledge is available about the eggs of laboratory animals, it will be possible to design more meaningful experiments in which the eggs of larger animals are used. In this way the most efficient use of embryos from the costly large domestic animals and primates can be made.

Until we have more information, a general medium for the cultivation of mammalian ova might consist of the following: (1) the salts contained in blood plasma, or Krebs-Ringer bicarbonate, (2) a sodium bicarbonate buffer with a concentration of approximately 25 mMoles, (3) an atmosphere containing 5% CO₂ in the gas phase, (4) a pH of approximately 7.4 which would be the result of items two and three, (5) a protein concentration of 1 to 10 mg./ml., (6) a glucose concentration of 1 mg./ml., (7) a pyruvate concentration of about 5 X 10_–4M. Supplementary compounds could be added to such a preparation, such as serum, embryo extracts, lactate, amino acids, vitamins and cofactors.

¹ The author would like to thank Dr. Joan Thomson for reading and commenting on the manuscript and Mrs. Peggy Earnest for the typing. Financial support for some of the research reported in this chapter has come from the National Science Foundation(GB 4465),the Population Council, and the Pennsylvania Department of Agriculture.

² King Ranch Laboratory of Reproductive Physiology. Department of Animal Biology, School of Veterinary Medicine.