Identification of Niche Conditions Supporting Short-term Culture of Spermatogonial Stem Cells Derived from Porcine Neonatal Testis

초록

Despite that porcine spermatogonial stem cells (pSSCs) have been regarded as a practical tool for preservingeternally genetic backgrounds derived from pigs with high performance in the economic traits or phenotypes ofspecific human diseases, there were no reports about precise definition of niche conditions promoting proliferation andmaintenance of pSSCs. Accordingly, we tried to determine niche conditions supporting proliferation and maintenanceof undifferentiated pSSCs for short-term. For these, undifferentiated pSSCs were progressively cultured in differentcomposition of culture medium, seeding density of pSSCs, type of feeder cells and concentration of growth factors,and then total number of and alkaline phosphatase (AP) activity of pSSCs were investigated at post-6 day culture. As the results, the culture of 4 × 105 pSSCs on mitotically in activated 2 × 105 STO cells in the mouse embryonicstem cell culture medium (mESCCM) supplemented with 30 ng/ml glial cell line-derived neurotrophic factor (GDNF)was identified as the best niche condition supporting effectively the short-term maintenance of undifferentiated pSSCs. Moreover, the optimized short-term culture system will be a basis for developing long-term culture system of pSSCsin the following researches.

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