Colonization by gene-corrected GS cells following testicular transplantation.

Abstract

<p>(A) Whole mounted squash preparation of seminiferous tubules depicting a seminiferous tubule (arrows) extensively colonized by gene-corrected GT59 SSCs two months following transplantation into a <i>Kit^W-v</i>/<i>Kit^W</i> sterile pup testis. (B) Non-transplanted control. (C) Whole mounted squash preparation of seminiferous tubules depicting a colony of GT65 SSCs two months following transplantation into a busulfan treated adult testis. Visualization of GT65 cells was facilitated by modification with Histone-GFP lentivirus prior to transplant. (A–C) Large arrowheads indicate GFP+ colonies and small arrows indicate autofluorescence in nearby tubules. Bar  = 100 microns. (C′) Higher magnification image of the boxed area in (C). Bar  = 50 microns. (D–G) Immunostaining with anti-GFP antibody (E, G, E′) or DAPI staining (D, F, D′, F′) of a cryosection of a GT59 colony 6 months following transplantation into <i>Kit^W-v</i>/<i>Kit^W</i> pup testis (D, E, D′, E′) or non-transplanted control testes (F, G, F′). Boxed area in D corresponds to the higher magnification view in D′ and E′. Triangles indicate donor-derived GFP+ cells. Boxed area in F corresponds to the higher magnification view in F′ and depicts the “Sertoli Cell Only” phenotype of non-transplanted <i>Kit^W-v</i>/<i>Kit^W</i> testes. The Sertoli cells are indicated by open triangles. Bar  = 25 microns.</p