By Michael S Kallos

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Establishment of hESC line on HFF and MEF feeder cells. Phase contrast microscopy of NS-10 line at different stages of development. A) The formation of outgrowth from the inner cell mass of blastocyst after direct culture of zona-free blastocyst on HFF. B) Cell clump, which was formed after mechanically dissociation of outgrowth, included primary hESC like cells. C) Circular primary NS-10 colony on MEF. D) Polarized colony morphology of NS-10 on HFF. 1% insulin transferrin selenium complex (Gibco BRL).

The sperm usually enters an egg a few hours after insemination. If there is a low chance for fertilization, one single sperm can be injected into an egg in a procedure called Intracytoplasmic Sperm Injection (ICSI). 4) Embryo culture. The fertilized eggs remain in the petri dish for 48 to 72 hours to verify that the embryo is not defective and growing properly. If a couple is at high-risk for passing on genetic (hereditary) disorders to a child they may consider using Pre-implantation Genetic Diagnosis (PGD).

19 The embryo thawing process is quite complex. “Embryo survival is based on the number of viable cells in an embryo after thawing. An embryo has ‘survived’ if >50% of the cells are viable. An embryo is considered to ‘partially survive’ if <50% of its cells are viable and to be ‘atretic’ if all the cells are dead at thaw. Approximately, 65-70% of embryos survive thaw, 10% partially survive and 20-25% are atretic. Data suggests that embryos with 100% cell survival are almost as good as embryos never frozen but only about 30-35% survive this fashion.

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