Abstract
The present study aimed to: (i) identify the exogenous factors that allow in vitro differentiation of mouse spermatogonial stem cells (SSCs) from embryonic stem cells (ESCs); (ii) evaluate the effects of Sertoli cells in SSC enrichment; and (iii) assess the success of transplantation using in vitro differentiated SSCs in a mouse busulfan-treated azoospermia model. A 1-day-old embryoid body (EB) received 5 ng/ml of bone morphogenetic protein 4 (BMP4) for 4 days, 3 µM retinoic acid (RA) in a SIM mouse embryo-derived thioguanine and ouabain resistant (STO) co-culture system for 7 days, and was subsequently co-cultured for 2 days with Sertoli cells in the presence or absence of a leukaemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and RA composition, and in the presence of these factors in simple culture medium. Higher viability, proliferation and germ cell gene expression were seen in the presence of the LIF, bFGF and RA composition, on top of Sertoli cells. Immunocytochemistry results showed higher CDH1 expression in this group. Sertoli co-culture had no effects on SSC proliferation. Eight weeks after transplantation, injected cells were observed at the base of the seminiferous tubules and in the recipient testes. The number of spermatogonia and the mass of the testes were higher in transplanted testes relative to the control group. It seems that transplantation of these cells can be useful in infertility treatment.
- basic fibroblast growth factor (bFGF)
- leukaemia inhibitory factor (LIF)
- retinoic acid
- transplantation
Abbreviations
- bFGF,
- basic fibroblast growth factor;
- BMP,
- bone morphogenetic protein;
- CDH1,
- cadherin 1;
- DMEM,
- Dulbecco’s modified Eagle’s medium;
- dpc,
- days postcoitus;
- EB,
- embryoid body;
- ESC,
- embryonic stem cell;
- GDNF,
- glial cell line derived neurotrophic factor;
- KSOM,
- potassium-rich medium derived by the simplex optimization method;
- LIF,
- leukaemia inhibitory factor;
- Mvh,
- mouse vasa homologue;
- PGC,
- primordial germ cell;
- Piwil2,
- piwi (Drosophila)-like 2;
- RA,
- retinoic acid;
- SCF,
- stem cell factor;
- SF,
- steel factor;
- SSC,
- spermatogonial stem cell;
- STO,
- SIM mouse embryo-derived thioguanine and ouabain resistant;
- Stra8,
- stimulated by retinoic acid 8;
- THY-1,
- thymus cell surface antigen 1
- © 2017 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society