Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources duringin vitroculture
| Autor: | A.L.P. Souza, A. W. B. Silva, B.R. Silva, L.R.F.M. Paulino, P A A Barroso, L B Aragão, D S Gomes, J. R. V. Silva, G.L. Vasconcelos, M F Lima Neto |
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| Rok vydání: | 2019 |
| Předmět: |
Granulosa Cells
biology Proteins Serum Albumin Bovine Histology Cell Biology In vitro Culture Media In Vitro Oocyte Maturation Techniques Andrology chemistry.chemical_compound Ovarian Follicle chemistry Follicular phase Oocytes biology.protein Animals Cattle Female Ethidium homodimer assay Basal membrane Bovine serum albumin Paraformaldehyde Fetal bovine serum Developmental Biology |
| Zdroj: | Zygote. 28:32-36 |
| ISSN: | 1469-8730 0967-1994 |
| Popis: | SummaryThe present study evaluated the effect of knockout serum replacement (KSR), fetal bovine serum (FBS) and bovine serum albumin (BSA) on the viability and growth of bovine secondary follicles culturedin vitrofor 12 days. To this end, secondary follicles were isolated (185–202 μm) and culturedin vitroin TCM-199+medium supplemented with KSR (5% and 10%), FBS (5% and 10%) or BSA (3 mg/ml) at 38.5°C with 5% CO2in air. Follicular diameters were evaluated on days 0, 4, 8 and 12. After 12 days of culture, follicular survival analysis was performing by using calcein-AM and ethidium homodimer. Before and after culture, follicles were fixed in paraformaldehyde for histological evaluation. Follicular diameter at different days of culture were compared using the Kruskal–Wallis test, while the percentages of viable follicles were analyzed by chi-squared test (P< 0.05). Results showed that follicles cultured in the presence of KSR at both concentrations presented higher follicular survival rates than those cultured in control medium alone or supplemented with FBS or BSA. Conversely, the presence of KSR, BSA or FBS did not increase follicular diameter after 12 days of culture. Histology analysis showed that, among the tested treatments, follicles cultured in the presence of KSR had preserved rounded oocytes, juxtaposed granulosa cells and intact basal membrane. In conclusion, supplementation of culture medium with KSR increases the follicular survival of bovine secondary follicles culturedin vitro. |
| Databáze: | OpenAIRE |
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