Cryopreservation of Spermatogonial Stem Cells of Native Goat of Iran

Subject Areas : Camel

M. Mohebbi ¹ , G. Moghaddam ² , B. Qasemi-Panahi ³ , H. Daghigh Kia ⁴ , S.A. Rafat ⁵ , G. Hamidian ⁶

1 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
2 - Department of Histology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran
3 - Department of Histology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran
4 - Department of Histology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran
5 - Department of Histology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran
6 - Department of Histology, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran

Received: 2019-07-08 Accepted : 2020-01-30 Published : 2021-06-01

Keywords: freezing, cryopreservation, Goat, Thawing, Co-culture, Sertoli cells, fetal bovine serum, cell isolation, spermatogonial cells,

Abstract :

This study deals with cryopreservation of spermatogonial stem cells (SSCs) of native goat using different media along with cryoprotectant. Morphological differentiation and immunocytochemistry tests were used to identify the cells. Furthermore anti-vimentin and anti-Oct-4 immuno staining methods were used for identification of sertoli cells and SSCs, respectively. Cryopreservation of SSCs was done with two sets of media. One with 40% dulbecco’s modified eagle’s medium (DMEM), 50% fetal bovine serum (FBS) and 10% dimethyl sulfoxide (DMSO). The second medium included 90% FBS and 10% DMSO. Post thaw cryopreserved cells were subjected to viability and colony area formation on 4th, 8th and 12th days of culturing after and before cryopreservation. Results clearly indicated the viability, area and number of colonies in the first medium registered to 61.37%, 0.87 mm2and 246.88 averagely in every 25 cm2 culture flask, respectively. Similarly with second medium, post thaw cryopreserved sperm registered viability, area and number of colonies to 73.87%, 2.74 mm2 and 364.36 in every 25 cm2 culture flask, respectively. After the thawing of spermatogonial cells, the best viability percentage was obtained in the freezing medium containing 90% FBS. Thus the study demonstrated that serum concentration had a distinct positive effect on the maintenance and proliferation of SSCs in culture after cryopreservation.

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