Ethics code: IR.RAZI.REC.1400.051
Department of Clinical Sciences, Faculty of Veterinary Medicin, Razi University, Kermanshah, Iran , drp.rahimi@gmail.com
Abstract: (167 Views)
Background and Aim: Developting culture and cryopreservation methods for spermatogonial stem cells (SSCs) is the foundation for genetic studies, regenerative medicine, production of transgenic animals and recombinant drugs, effort to stablish a suitable culture system for these cells is very important. The aim of this study was to investigate the in vitro effects of different concentrtions of L-Arginine on the proliferation of frozen-thawed caprine SSCs.
Methods: The present study is experimental and was repeated 5 times.
The isolated spermatogonial cells were frozen-thawed and cultured in six groups, they were added to basal medium culture (DMEM containing 1% antibiotics and 5% FBS) containing different concentrations of L-arginine (0, 50, 100, 200, 500, and 1000 micromoles per liter of L-arginine), respectively. All experimental groups were cultured for 10 days. The number and surface area of colonies formed were evaluated on days 4, 7 and 10. Expression of spermatogonial genes (ID4, BCL6, PGP9.5, VASA, PLZF, OCT4) in the culture was determined by RT-PCR.
Results: The viability rates of fresh cells before and after the addition of cryoprotectant agent were 87.14±2.6 and 80.72±1.21, respectively which decreased to 65.38±1.26 after the thawing. The number and surface area of spermatogonia colonies in the fourth treatment (200 μmol / l L-arginine) was statistically significantin camparision with other experimental groups (P<0.05). Finally, the isolated cells expressed SSCs surface markers.
Conclusion: Probably, adding of 200 μmol / l L-arginine has a desirable effect on the induction of goat frozen-thawed SSCs colonization in short-term culture.
Type of Study:
Original |
Subject:
Cellular Sciences (Molecular Cells, Stem Cells) Received: 2024/05/25 | Accepted: 2024/12/23 | Published: 2025/03/11
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