Embryo Project Encyclopedia Articles

In 1972, Peter Mazur, Stanley Leibo, and Ernest Chu published, “A Two-Factor Hypothesis of Freezing Injury: Evidence from Chinese Hamster Tissue-culture Cells,” hereafter, “A Two-Factor Hypothesis of Freezing Injury,” in the journal, Experimental Cell Research. In the article, the authors uncover that exposure to high salt concentrations and the formation of ice crystals within cells are two factors that can harm cells during cryopreservation. Cryopreservation is the freezing of cells to preserve them for storage, study, or later use. Mazur originally suggested the two factors in a 1970 paper, but that article was based on evidence from simple yeast cells. By using hamster cells in 1972, Mazur, Leibo, and Chu confirmed that Mazur’s two-factor hypothesis applied to more complex mammalian cells. The article dispelled the widely accepted notion that rapid cooling rates were safest for all cells, and instead showed that each kind of cell had a different optimal cooling rate depending on the solution in which it froze.

Stanley Paul Leibo studied the cryopreservation of embryos in the US in the twentieth century. Cryopreservation is a method of preserving biological material through freezing. Early in his career, Leibo collaborated with other scientists to study why cells were oftentimes injured during freezing. Later, Leibo and his team accomplished one of the first successful births using previously-frozen mammalian embryos. Leibo continued evolving simpler and more reliable methods of cryopreservation and embryo transfer for many different species over the course of his career, such as the development of a one-step procedure of transferring fertilized embryos between cattle. Leibo’s work to develop simple and reliable ways to cryopreserve cells and embryos enabled its use in a wider scope of research, including agriculture, reproductive medicine, and conservation.