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Bovine Sertoli Cells Colonize and Form Tubules in Murine Hosts Following Transplantation and Grafting Procedures

ZHEN ZHANG^*,

JON HILL

MICHAEL HOLLAND^*,

YASUYUKI KURIHARA

KATE L. LOVELAND^*,

From the ^* Center for Reproduction and Development, Monash Institute of Medical Research, Monash University, Clayton, Australia; CSIRO Livestock Industries, Armidale, Australia; Australian Research Council Centre of Excellence in Biotechnology and Development, Australia; and Department, of Environment and Natural Sciences, Graduate School of Environment and Information Sciences, Yokohama National University, Yokohama, Japan.

Correspondence to: Dr Zhen Zhang, Center for Reproduction and Development, Monash Institute of Medical Research, Monash University, 27–31 Wright St Clayton, VIC 3168, Australia (e-mail: zhen.zhang{at}med.monash.edu.au).

The contribution of somatic cells to nonrodent male germ cell transplantation success has not been well established due to lack of cell type-specific markers to distinguish donor cells from host cells. In the present study, we first screened antibodies and a lectin to identify markers suitable for unequivocal distinction between germ cells and Sertoli cells in bovine testes compared with mouse testes. Anti-vimentin and the Dolichos biflorus agglutinin (DBA) lectin detected only bovine Sertoli cells and spermatogonia, respectively; anti-NONO and anti-GCNA1 detected only mouse Sertoli and germ cells, respectively. The outcome of transplanting bovine testis cells into nude mouse testes was then studied using these markers. Our results clearly showed that immature bovine Sertoli cells survive and colonize mouse testes at 2.5 months after transplantation and that tubular structures composed of donor Sertoli cells formed adjacent to murine tubules within the host mouse testis. Bovine germ cell colonization and survival in mouse testes after transplantation were confirmed, but this was restricted to areas of bovine Sertoli cell colonization. In addition, ectopic grafts of intact bovine testis tissue and cell aggregates from hanging drop cultures were placed under the back skin and testis capsule of nude mice. Bovine Sertoli cells in ectopic grafts and aggregates were able to form tubular structures, and some bovine germ cells were observed around 2 months after implantation. This study therefore identifies a practical strategy to assess the outcome of testicular cell transplantation using different antibodies and a lectin to distinguish bovine cells from mouse cells. It identifies an approach that can readily be adapted to study other nonrodent species.

     Key words: Bovine spermatogonia, cell aggregates, xenotransplantation, antibodies

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