earticle

영어

Spermatogonial stem cells (SSC) are promising resources for genetic preservation and restoration of male germ cells in human and animals. However, in our preliminary experiment, it was observed that SSC was fragile when used as nuclei donor for SCNT. This study investigated the potential of porcine SSC as donor nuclei for somatic cell nuclear transfer (SCNT) and developmental competence of SSC-derived cloned embryos. In addition, it was examined whether demecolcine could prevent rupture of SSC during SCNT. SCNT embryos were produced using a standard protocol of our laboratory. After electric activation, SCNT embryos were treated with demecolcine combined with 6-DMAP for 4 h, washed, and then cultured in a porcine zygote medium-3 at 39℃ in a humidified atmosphere of 5% CO2, 5% O2, and 90% N2 for 7 days. When SSC was compared with porcine fetal fibroblast (PFF) in the potential to support embryonic development after SCNT, SSC-derived SCNT embryos showed higher (p< 0.05) developmental competence to the blastocyst stage (53.7%) than PFF-derived embryos (28.6%). Treatment of SSC with demecolcine significantly (p<0.01) inhibited the rupture of SSC during SCNT (7.8% vs. 16.4%) and increased fusion of cell-ooplasm couplets compared to no treatment (71.9% vs. 62.8%). In addition, even after demecolcine treatment, SSC-derived SCNT embryos showed a higher blastocyst formation (46.4%) than PFF-derived embryos (27.8%). Our results demonstrate that porcine SSC are a desirable donor cell type for production of SCNT pig embryos and demecolcine increases production efficiency of cloned embryos by inhibiting rupture of nuclei donor SSC during SCNT.