Proliferation and Differentiation of Mouse Spermatogonial Stem Cells on a Three-Dimensional Surface Composed of PCL/Gel Nanofibers

Abstract

Spermatogonial stem cells (SSCs) have self-renewal and differentiation capacity essential for sperm production throughout the male reproductive life. The electrospun polycaprolactone/gelatin (PCL/Gel) nanofibrous scaffold mimics important features of the extracellular matrix (ECM), which can provide a promising technique for the proliferation and differentiation of SSCs in vitro. The goal of the present study was to investigate the effects of PCL/Gel nanofibrous scaffold on the propagation and differentiation of neonate mouse SSCs (mSSCs). mSSCs were enzymatically isolated, and the cells were purified by differential plating method and seeded on scaffold. After 2 weeks, viability, colony number and diameter, and expression of specific spermatogonial cell genes were investigated. After mSSCs propagation, the cells were cultivated in a differentiation medium on the scaffold for another 2 weeks, and differentiating cells were analyzed by real-time PCR. The number of mSSC colony (P<0.01) and expression levels of specific spermatogonial genes Plzf and Inga6 (P<0.01) and also differentiation genes c-Kit, Tp1 and Ptm1 (P<0.05) were higher in scaffold group compared with control during the culture period. We conclude that mSSCs can be expanded and can differentiate toward spermatid cells on PCL/Gel nanofibrous scaffold with improved developmental parameters.

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

“…Liao et al have also used an irregular electrospun nanoparticle, in order to fabricate a network for SSCs culturing, and as a result, they have indicated that this structure resembled a basement membrane and also provided a suitable environment for the cell growth (Liao et al, 2006). Furthermore, Talebi et al have used a PCL–Gel scaffold to culture mouse spermatogonia and then have reported that the scaffold is able to maintain stem cell's viability and proliferation (Talebi et al, 2019).…”

“…In this regard, some studies showed that SCF/c‐kit system is very important for regulating survival, proliferation, apoptosis, differentiation and onset of meiosis in SSCs. As well, defects in SCF/c‐kit system of testis could lead to cessation of differentiation in these cells and to infertility (Lennartsson & Rönnstrand, 2012; Talebi et al, 2019).…”

Biomaterials for Testicular Bioengineering: How far have we come and where do we have to go?

“…The use of synthetic polymers in tissue bioengineering is indicated because such polymers are biocompatible, have suitable physical-mechanical properties and elasticity to produce scaffolds. Among the synthetic materials used for the composition of scaffolds, there are the poly (L-lactic acid) (PLLA) ( 195 , 213 , 214 ), polypropylene, poly D, polycaprolactone (PCL) ( 215 ) and L-lactic-co-glycolic acid ( 208 ), which have already been used in the production of bone ( 227 ), cartilaginous ( 228 ), vascular ( 229 ) and dermal ( 230 ) scaffolds. Synthetic polymers can be combined with natural polymers, which improves their mechanical, physical and biocompatibility qualities.…”

Advances of three-dimensional (3D) culture systems for in vitro spermatogenesis

“…Survival rate of SSCs cultured on PCL/Gelatin nanofibrous scaffolds was significantly higher than control group. Talebi et al [ 148 ] Neonatal mouse SSCs Electrospun nanofibrous PCL/Gel scaffold 4 weeks Number of colonies and viability rate of cultured cells on PCL/Gel scaffold increased. Expression of spermatogonial markers (Plzf and Inga6), meiotic and post meiotic genes (c-Kit, Tp1, and Prm1) significantly increased compared to the control group.…”