Cloned embryos from semen. Part 1: In vitro proliferation of epithelial cells on embryonic fibroblasts after isolation from semen by gradient centrifugation

摘要: Although epithelial-like somatic cells have been previously isolated from semen, cell proliferation rates were low. Culture of whole semen samples resulted in loss potentially valuable spermatozoa. The aims the present study to: (1) isolate while preserving sperm viability, and (2) optimize vitro culture conditions for semen-derived epithelial cells. Density gradient centrifugation washed ejaculates two rams (Ovis aries) (n = 24) one eland bull (Taurotragus oryx) 4) was performed using a three-layer discontinuous Percoll column consisting 90% (P-90), 50% (P-50), 20% (P-20) Percoll. In Trypan Blue staining indicated that live settled P-20 layer. Nonmotile spermatozoa recovered at P-50 P-90 interfaces, whereas motile collected pellet Subsequently, layer plated either on inactivated 3T3 mouse embryonic fibrobla...