Effects of Extracellular Matrix Protein-derived Signaling on the Maintenance of the Undifferentiated State of Spermatogonial Stem Cells from Porcine Neonatal Testis

  • Park, Min Hee (Department of Animal Life Science, Kangwon National University) ;
  • Park, Ji Eun (Department of Animal Life Science, Kangwon National University) ;
  • Kim, Min Seong (Department of Animal Life Science, Kangwon National University) ;
  • Lee, Kwon Young (College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University) ;
  • Hwang, Jae Yeon (Division of Applied Animal Science, Kangwon National University) ;
  • Yun, Jung Im (Division of Animal Resource Science, Kangwon National University) ;
  • Choi, Jung Hoon (College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University) ;
  • Lee, Eunsong (College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University) ;
  • Lee, Seung Tae (Department of Animal Life Science, Kangwon National University)
  • Received : 2015.10.20
  • Accepted : 2016.01.25
  • Published : 2016.10.01


In general, the seminiferous tubule basement membrane (STBM), comprising laminin, collagen IV, perlecan, and entactin, plays an important role in self-renewal and spermatogenesis of spermatogonial stem cells (SSCs) in the testis. However, among the diverse extracellular matrix (ECM) proteins constituting the STBM, the mechanism by which each regulates SSC fate has yet to be revealed. Accordingly, we investigated the effects of various ECM proteins on the maintenance of the undifferentiated state of SSCs in pigs. First, an extracellular signaling-free culture system was optimized, and alkaline phosphatase (AP) activity and transcriptional regulation of SSC-specific genes were analyzed in porcine SSCs (pSSCs) cultured for 1, 3, and 5 days on non-, laminin- and collagen IV-coated Petri dishes in the optimized culture system. The microenvironment consisting of glial cell-derived neurotrophic factor (GDNF)-supplemented mouse embryonic stem cell culture medium (mESCCM) (GDNF-mESCCM) demonstrated the highest efficiency in the maintenance of AP activity. Moreover, under the established extracellular signaling-free microenvironment, effective maintenance of AP activity and SSC-specific gene expression was detected in pSSCs experiencing laminin-derived signaling. From these results, we believe that laminin can serve as an extracellular niche factor required for the in vitro maintenance of undifferentiated pSSCs in the establishment of the pSSC culture system.


Porcine;Spermatogonial Stem Cells;Undifferentiation;Extracellular Matrix Proteins


Supported by : Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET)


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