Effect of a Freezing Medium Containing Melatonin on Markers of Pre-meiotic and Post-meiotic Spermatogonial Stem Cells (SSCs) After Transplantation in an Azoospermia Mouse Model Due to Testicular Torsion

Spermatogonial stem cells (SSCs) are essential to the initiation of spermatogenesis. Cryopreservation, long-term maintenance, and auto-transplantation of SSCs could be a new treatment for infertility. The aim of this study was to add melatonin to the basic freezing medium and to evaluate its effect...

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Published in:Reproductive sciences (Thousand Oaks, Calif.) Calif.), 2021-05, Vol.28 (5), p.1508-1522
Main Authors: Kazemzadeh, Shokoofeh, Rastegar, Tayebeh, Zangi, Bagher Minaei, Malekzadeh, Mehrnoush, Khanehzad, Maryam, Khanlari, Parastoo, Madadi, Soheila, Bashghareh, Alieh, Hedayatpour, Azim
Format: Article
Language:English
Subjects:
Adult Germline Stem Cells - drug effects
Adult Germline Stem Cells - physiology
Adult Germline Stem Cells - transplantation
Animals
Azoospermia - complications
Azoospermia - prevention & control
Cells, Cultured
Cryopreservation - methods
Culture Media - pharmacology
Disease Models, Animal
Embryology
Male
Male Reproduction: Original Article
Medicine
Medicine & Public Health
Meiosis - drug effects
Melatonin - administration & dosage
Mice
Obstetrics/Perinatology/Midwifery
Reproductive Medicine
Spermatic Cord Torsion - complications
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Summary:Spermatogonial stem cells (SSCs) are essential to the initiation of spermatogenesis. Cryopreservation, long-term maintenance, and auto-transplantation of SSCs could be a new treatment for infertility. The aim of this study was to add melatonin to the basic freezing medium and to evaluate its effect on the efficiency of the thawed SSCs after transplantation into the testicles of azoospermic mice. SSCs were isolated from newborn NMRI mice, and the cells were enriched to assess morphological features. The thawed SSCs were evaluated for survival, apoptosis, and ROS level before transplantation, and the proliferation (MVH and ID4) and differentiation (c-Kit, SCP3, TP1, TP2, and Prm1) markers of SSCs were examined using immunofluorescence, western blot, and quantitative real-time polymerase chain reaction (PCR) after transplantation. It was found that the survival rate of SSCs after thawing was significantly higher in the melatonin group compared with the cryopreservation group containing basic freezing medium, and the rate of apoptosis and level of ROS production also decreased significantly in the cryopreservation group with melatonin ( p  
ISSN:1933-7191
1933-7205
DOI:10.1007/s43032-020-00447-5