221 Isolation of turkey spermatogonia
| Autor: | H. V. Ashraf, N. A. Zinovieva, Anastasia N Vetokh, N. A. Volkova, V. A. Bagirov, E. K. Tomgorova, L. A. Volkova |
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| Rok vydání: | 2020 |
| Předmět: |
endocrine system
Spermatogenic Cell education.field_of_study urogenital system Population Reproductive technology Biology Sertoli cell Cryopreservation Andrology Endocrinology medicine.anatomical_structure Reproductive Medicine Cell culture Genetics medicine Animal Science and Zoology education Molecular Biology Spermatogenesis Gametogenesis Developmental Biology Biotechnology |
| Zdroj: | Reproduction, Fertility and Development. 32:238 |
| ISSN: | 1031-3613 |
| DOI: | 10.1071/rdv32n2ab221 |
| Popis: | Across different types of spermatogenic epithelium, researchers have focused on the study and production of spermatogonial stem cells and their use as genetic material for creating gene pool cryobanks of valuable breeds and lines of poultry. The aim of our research was to identify optimal conditions for obtaining and culturing turkey spermatogonia. A series of experiments were carried out to determine the optimal age of males for isolating spermatogonia and optimize conditions for their isolation and cultivation. Age-related features of turkey spermatogenesis were assessed by testis histological studies. Statistical analysis was performed using SPSS v15.0 (t-test; SPSS Inc./IBM Corp.). Turkey testis tissue was mechanically crushed and treated with enzymes: 0.1% collagenase and 0.25% trypsin. Purification of spermatogonia from other types of cells was performed by adhesion and separation. The conditions and duration of spermatogenic cell cultures were selected experimentally. Primary Sertoli cells of turkeys, Sertoli cells of roosters, STO cell line, and transplanted Sertoli cells of pigs (PTP) were used as a feeder layer for the cultivation of spermatogonia. Spermatogonia culture medium consisted of Dulbecco's modified Eagle's medium (DMEM)-high glucose supplemented with 5% fetal calf serum, 2mM α-glutamine, 10µLmL−1 MEM, 100x antibiotics, 10µLmL−1 ITS solution, 5×10−5 M mercaptoethanol, 5mgmL−1 albumin, 1µLmL−1 DL-lactic acid, 20ngmL−1 epidermal growth factor, 10ngmL−1 basic fibroblast growth factor, and 2ngmL−1 leukemia inhibitory growth factor (LIF). The SSEA-1 antibody was used for identification of spermatogonia colonies. Histological study of the turkey testes at different ages showed that in animals under 12 weeks, the population of generative cells was represented mainly by spermatogonia (P |
| Databáze: | OpenAIRE |
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