Human spermatogonia stem cells (SSCS) in a culture system with platelet rich plasma and correlations with spermatogenesis level

Silvia W Lestari, Debby Aditya, Firda Asma’ul Husna, Gita Pratama, Dwi Anita Suryandari, Kanadi Sumapradja, Ria Margiana, Puji Sari, Ria Kodariah 

Department of Medical Biology, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia

ABSTRACT

There are instances when the sperm fail to be obtained from the testes in assisted reproductive technology (ART) of azoospermia infertility due to spermatogenesis arrest. Therefore, spermatogonial stem cells (SSCs) may be used as an alternative management. Platelet rich plasma (PRP) is a concentrated source of growth factors that support the proliferation and differentiation of stem cells in vitro. The objective of this study was to determine the efficiency of PRP in supporting SSCs proliferation and differentiation and to assess the correlation between the level of spermatogenesis and the potency of proliferation and differentiation of SSC in vitro. SSCs were isolated from seven testicular tissue samples using sperm extraction (TESA/TESE) and were then further cultured into two groups, PRP and FBS. The resulting cells were quantitatively assayed by qRT-PCR towards the expression of PLZF, OCT4, and CKIT. The level of spermatogenesis was assessed from the histology measurement using Johnson scoring. Apoptosis was evaluated using the TUNEL and immunocytochemistry methods. This study showed that PLZF, OCT4, and CKIT were expressed in the resulting cell culture. The difference was statistically insignificant among PRP and FBS. There was a positive correlation between the potency of proliferation and differentiation of SSCs toward the level of spermatogenesis. PRP may support the maintenance of proliferation and differentiation of SSCs in vitro and could be developed as an alternative of FBS in the SSCs culture system.

Keywords:

Spermatogonial stem cells, Platelet rich plasma, Fetal bovine serum, Spermatogenesis, Cell culture