Bovine ovarian stem cells differentiate into germ cells and oocyte-like structures after culture in vitro
| Autor: | R.P. Ribeiro, J. R. V. Silva, J.R.S. Passos, J.J.N. Costa, E. V. da Cunha, R. van den Hurk, G. B. de Souza, Márcia Viviane Alves Saraiva |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
endocrine system Cell Culture Techniques Biology Andrology 03 medical and health sciences Endocrinology medicine Animals Cells Cultured Stem Cells Ovary Cell Differentiation Amniotic stem cells Oocyte Follicular fluid Germ Cells 030104 developmental biology medicine.anatomical_structure P19 cell Amniotic epithelial cells embryonic structures Oocytes Cattle Female Animal Science and Zoology Stem cell Germ cell Biotechnology Adult stem cell |
| Zdroj: | Reproduction in Domestic Animals. 52:243-250 |
| ISSN: | 0936-6768 |
| Popis: | Stem cells have been isolated from ovaries, and their ability to differentiate into oocytes in vitro has been demonstrated for mice and human, but not for bovine species. The aims of this study were to isolate germline stem cells from bovine ovaries and to evaluate the effects of bone morphogenetic proteins (BMPs) 2 and 4, and follicular fluid on the differentiation of these stem cells into oocyte-like structures. The ovarian stem cells were isolated and cultured in α-MEM+ supplemented with BMP2, BMP4 or follicular fluid. On days 0 and 14, cells were evaluated for their morphological appearance, viability, expression of alkaline phosphatase and for markers of germ cell formation (VASA and DAZL) and oocyte development (GDF9, ZPA and SCP3) by qPCR. Levels of mRNA were analysed using ANOVA and Bonferroni test (p < .05). The results showed that at day 0, ovarian stem cells expressed specific markers of pluripotency (OCT4, SOX). In addition, these cells were positive for alkaline phosphatase, which is a marker commonly used to identify primordial germ cells (PGCs). After the period of differentiation, cells had morphological features that resemble PGCs and oocyte-like cells (OLCs). An increase, ranging from five to 14 times, in the expression of VASA was observed in cells cultured in medium supplemented with BMPs and follicular fluid, while the increase in DAZL expression ranged from four to six times. In addition, OLCs had an increase in expression of mRNAs for GDF9, ZPA and SCP3 that ranged from two to eight times. In conclusion, OLCs can be differentiated in vitro from ovarian stem cells and BMPs and follicular fluid are effective in stimulating the expression of mRNAs for germ cell and oocyte markers. |
| Databáze: | OpenAIRE |
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