Long-term culture and transplantation of XXXX testicular germ cells.

@entity25325 @entity1603 @entity28895 @entity19 @entity9640 @entity17777 @entity21923 @entity1778 @entity6514 @entity28842 @entity2241
The objectives of this study were to develop an in vitro culture system to optimize germ cell proliferation and to measure the potential of the cultured germ cells to produce mature spermatozoa after transplantation into a recipient. Donor germ cells isolated from @entity25325 male @entity19 were cultured with a STO feeder cell layer in Dulbecco's minimal essential medium (DMEM) supplemented with @entity2241 ( @entity2241 ), stem cell factor, @entity1778 inhibitory factor, basic fibroblast growth factor, insulin-like growth factor 1, @entity28895 , @entity9640 , @entity21923 , @entity6514 , @entity19 @entity28842 , and platelet-derived growth factor. Donor germ cells formed colonies in the primary cultures after 8-21 days. These cultured colonies were maintained for 4 weeks or longer without subculture and proliferated for up to 8 passages over a period of 3 months. These colonies had alkaline phosphatase activity and incorporated @entity1603 . These colonies were positive partially when screened with antibody for germ cell nuclear antigen and @entity17777 . Germ cells cultured with this supplemented medium showed enhanced colonization vs controls cultured with DMEM and @entity2241 . Cultured germ cells from @entity25325 donors were transplanted into testes and were identified by X-gal staining and histological screening. The cells cultured in the supplemented medium colonized the tubules and initiated spermatogenesis in the recipient @entity19 . This is an improved method for culturing germ cells and may be useful in gene therapy and the production of transgenic animals.
