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

Abstract

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.

Proliferation of human spermatogonial stem cells on optimized PCL/Gelatin nanofibrous scaffolds

“…It has also been shown that integrins α6 and β1 are expressed on the surface of spermatogonia stem cells. Accordingly, these could also bind cells to laminin in the basement membrane of seminiferous tubules (Park et al, 2016). Besides spermatogenic stem cells, integrin α6 (CD49) was also indicated to be expressed in Sertoli cells (Schlatt, 2002).…”

Distribution of extracellular matrix related proteins in normal and cryptorchid ziwuling black goat testes

“…ECM components of testes are mainly produced by Sertoli cells in seminiferous tubules and peritubular myoid cells in the lamina propria, both of which cooperate with each other in the formation of ECM. Generally, Sertoli cells generate Col IV, LN, actin and HSPG, while the peritubular myoid cells synthesizes LN, ColI, and Col IV ( Park et al, 2016 ). Studies have shown that the important components of testicular ECM, such as Col IV, LN and HSPG, interact with proteases and protease inhibitors of spermatogenic cells, TNFα and other cytokines to mutually regulate spermatogenesis ( Siu and Cheng, 2008 ).…”

“…LN is widespread in the matrix, and it is the most abundant ECM component in the basement membrane, which has many linking and mediating effects to controll the formation of basement membrane ( Binsila et al, 2018 ; Park et al, 2016 ). Cooperation between Sertoli cells and peritubular myoid cells contribute to the secretion of LN.…”