44 EFFECT OF CRYOPROTECTANT AGENTS IN EQUINE OVARIAN BIOPSY FRAGMENTS
| Autor: | G.M. Ishak, S. T. Bashir, G. D. A. Gastal, S. O. Paiva, Benner Geraldo Alves, S. G. S. de Tarso, Eduardo L. Gastal, K.A. Alves |
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| Rok vydání: | 2016 |
| Předmět: |
Stromal cell
Cryoprotectant Dimethyl sulfoxide Reproductive technology Biology Oogenesis Cryopreservation Andrology chemistry.chemical_compound Endocrinology Reproductive Medicine chemistry Immunology Genetics Animal Science and Zoology Folliculogenesis Molecular Biology Ethylene glycol Developmental Biology Biotechnology |
| Zdroj: | Reproduction, Fertility and Development. 28:152 |
| ISSN: | 1031-3613 |
| Popis: | Studies on ovarian tissue preservation in horses are scarce, and only one research group (Devireddy et al. 2006 Mol. Reprod. Develop. 73, 330–341) has tested the effect of the cryoprotectant agents in equine ovarian tissue. The purpose of this study was to evaluate (1) the effect of cryoprotectant agents in the morphology of equine preantral follicles (PAF) and structure of the ovarian tissue, (2) the relationship between ovarian stromal cell density and PAF density, and (3) the association of ovarian stromal cell density with PAF morphology after exposure to cryoprotectant agents. Three independent experiments with identical methodologies were performed in this study. Each experiment was composed by one cryoprotectant agent (dimethyl sulfoxide, ethylene glycol, propylene glycol) performed in 3 replicates. Ovarian biopsy fragments (1.5 × 1.5 × 10 mm) were harvested from 6 mares and submitted to 4 times of exposure (0, 10, 15, 20 min) at room temperature in a solution of α-minimal essential medium (MEM)+ and one cryoprotectant (dimethyl sulfoxide, ethylene glycol, or propylene glycol) at a concentration of 1.5 M. The cryoprotectant removal was performed by a 3-step (5 min each) equilibration process using (1) α-MEM + 0.5 M sucrose, (2) α-MEM + 0.25 M sucrose, and (3) α-MEM alone after each time of exposure. All fragments were fixed in Bouin’s solution for histological analysis. Data were analysed by ANOVA, Fisher’s exact test, Spearman’s rank correlation, and power test. The PAF density, stromal cell density, and area of the fragments were not affected (P > 0.05) by any of the cryoprotectant agents throughout the time of exposure. However, the morphology of the PAF was affected (P 0.05) throughout the times of exposure. Strong correlations (range, 0.57 to 0.77; power, 96 to 99%) were identified between PAF density and stromal cell density in all experiments. When stromal cells were classified into groups of density (i.e. low, 25.67–34.66; medium, 34.67–43.66; and high, 43.67–52.67 cells/2500 μm2), only the high stromal cell density was positively correlated (P |
| Databáze: | OpenAIRE |
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