Encyclopedia of Reproductive Technologies

By Annette Burfoot | Go to book overview
identify. Tests in which cell proliferation is measured (in mouse embryos and other types of cell culture) are very sensitive and may be able to detect subtle toxicity problems.The purity of the water used in the preparation of culture media is extremely important. The source of water varies widely, including rainwater distilled multiple times and city water treated by reverse osmosis and multiple distillations. The most commonly used method for water purification is reverse osmosis followed by ultrafiltration and passage through a pyrogen filter. Considerable care is required to maintain ultrafiltration systems, and routine maintenance is critical.All other aspects of the IVF laboratory require strict quality control. Incubators must be routinely cleaned and sterilized. Temperature and carbon dioxide levels must be closely monitored. Adequate humidification is essential. All glassware in contact with culture media must be washed and sterilized using the tissue culture method. Washing procedures should include soaking in dilute acid solution, sonication, and thorough rinsing in ultrapure water. When detergents are used, they must be completely removed by proper rinsing procedures. Dry-heat sterilization is commonly used for human IVF. Ethylene oxide can also be used, but adequate venting is necessary to remove all traces of gas. Autoclaving is not recommended, and irradiation should not be used on some plastics. Sterile technique is essential to avoid contamination of the culture system with microorganisms. Yeast and fungi are common contaminants known to have detrimental effects on gametes and embryos.
Conclusion
Fertilization and early embryo development occur in the fallopian tube, and culture systems for IVF have attempted to mimic this environment. However, due to the inaccessibility of reproductive tract fluids for analysis and the dynamic nature of the tubal environment, only moderate success has been achieved. Nevertheless, human IVF has had a remarkable history and has experienced incredible growth since the birth of the first IVF baby in 1978. In vitro fertilization has become an accepted therapeutic procedure for coping with many types of infertility and is continuing to broaden in scope. Future developments will depend on media formulations and culture conditions that have been improved to meet the metabolic requirements of eggs and embryos.
FOR FURTHER READING
Bongso, A., et al. 1995. "Coculture Techniques for Embryonic Stem Cell Production." Assisted Reproduction Reviews 5: 106-114.

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