Volume 48, Issue 4 (2-2025)
Research in Medicine 2025, 48(4): 14-25 |
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Ethics code: IR.RAZI.REC.1400.051
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Najafi F, Rahimi-Feyli P, Moghaddam A, Mohammadi T. The Effect of L-Arginine on Growth of Frozen- Thawed Caprine Spermatogonial Stem Cells in Short- Term Culture. Research in Medicine 2025; 48 (4) :14-25
URL: http://pejouhesh.sbmu.ac.ir/article-1-3380-en.html
URL: http://pejouhesh.sbmu.ac.ir/article-1-3380-en.html
Department of Clinical Sciences, Faculty of Veterinary Medicine, Razi University, Kermanshah, Iran. , drp.rahimi@gmail.com
Abstract: (437 Views)
Background and Aim: Developing culture and cryopreservation methods for spermatogonial stem cells (SSCs) is essential for genetic studies, regenerative medicine, transgenic animal production and recombinant drug development. Establishing an optimal culture system for these cells is crucial. This study aimed to investigate the in vitro effects of different concentrations of L-arginine on the proliferation of frozen-thawed caprine SSCs.
Methods: This experimental study was repeated five times. Isolated spermatogonial cells were frozen- thawed and cultured in six groups with a basal culture medium (DMEM containing 1% antibiotics and 5% FBS) supplemented with different concentrations of L-arginine (0, 50, 100, 200, 500 and 1000 μmol/L). All experimental groups were cultured for 10 days. The number and surface area of colonies formed were assessed on days 4, 7, and 10. Expression of spermatogonial genes (ID4, BCL6, PGP9.5, VASA, PLZF and OCT4) was determined by RT-PCR.
Results: The viability rates of fresh cells before and after the addition of a cryoprotectant agent were 87.14 ± 2.6% and 80.72 ± 1.21%, respectively, which decreased to 65.38 ± 1.26% after thawing. The number and surface area of spermatogonial colonies in the fourth treatment group (200 μmol/L L-arginine) were significantly higher compared to other experimental groups (P < 0.05). Finally, the isolated cells expressed SSC surface markers.
Conclusion: The addition of 200 μmol/L L-arginine likely enhances the colonization of frozen-thawed goat SSCs in short- term culture.
Methods: This experimental study was repeated five times. Isolated spermatogonial cells were frozen- thawed and cultured in six groups with a basal culture medium (DMEM containing 1% antibiotics and 5% FBS) supplemented with different concentrations of L-arginine (0, 50, 100, 200, 500 and 1000 μmol/L). All experimental groups were cultured for 10 days. The number and surface area of colonies formed were assessed on days 4, 7, and 10. Expression of spermatogonial genes (ID4, BCL6, PGP9.5, VASA, PLZF and OCT4) was determined by RT-PCR.
Results: The viability rates of fresh cells before and after the addition of a cryoprotectant agent were 87.14 ± 2.6% and 80.72 ± 1.21%, respectively, which decreased to 65.38 ± 1.26% after thawing. The number and surface area of spermatogonial colonies in the fourth treatment group (200 μmol/L L-arginine) were significantly higher compared to other experimental groups (P < 0.05). Finally, the isolated cells expressed SSC surface markers.
Conclusion: The addition of 200 μmol/L L-arginine likely enhances the colonization of frozen-thawed goat SSCs 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
Received: 2024/05/25 | Accepted: 2024/12/23 | Published: 2025/03/11
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