Abstract
The aim of this study was to evaluate isolated bovine preantral follicles after vitrification procedure. Ovaries were obtained at a local slaughterhouse and submitted to follicular isolation. From the obtained suspension, one part was destined to control group. The remaining suspension was divided into two parts, one to the toxicity test and the other one to the vitrification procedure. For vitrification, the follicles were exposed for 1 min to vitrification solution-1, composed of 10% ethylene glycol, 10% DMSO in saline solution (NaCl 0.9%), and then 1 min to vitrification solution-2, composed of 20% ethylene glycol, 20% DMSO in the same saline solution supplemented with 0.5 M sucrose. The follicles were vitrified by use of glass capillaries and remained vitrified for 24 h. After warming, they were transferred to a 0.25 M sucrose solution for 5 min and then transferred to a 0.15 M sucrose solution for another 5 min. For the toxicity test, the follicles were only exposed to cryoprotectants and analised. The preantral follicles were classified as non-viable orviable when they were stained or not stained by trypan blue, respectively. Differences (P<0.05) in follicular viability were observed among the control, non-vitrified group (93.6%), and exposed (64.1%) or vitrified follicles (66%). The percentages of normal follicles found after exposure to cryoprotectant did not differ significantly (P>0.05) from that found after vitrification, indicating that the reduction of percentage of viable preantral follicles was due to a toxic effect of cryoprotectants and not to the vitrification procedure itself.KEYWORDS: bovine; cryoprotectants; oocytes; vitrification.
Highlights
Observou-se diferença (93,6%) do Grupo 1 com os Grupos 2 (64,1%) e 3 (66%), mas não entre os dois últimos
After exposure to cryoprotectant did not differ significantly was due to a toxic effect of cryoprotectants and not to the (P>0.05) from that found after vitrification, indicating that vitrification procedure itself. the reduction of percentage of viable preantral follicles
CELESTINO, J.J.H.; SANTOS, R.R.; LOPES, C.A.P.; MARTINS, F.S.; MATOS, M.H.T.; MELO, M.A.P.; BÁO, S.N.; RODRIGUES, A.P.R.; JOSÉ ROBERTO VIANA SILVA, J.R.V.; FIGUEIREDO, J.R. Preservation of bovine preantral follicle viability and ultra-structure after cooling and freezing of ovarian tissue
Summary
Os ovários (n=5) foram coletados, em abatedouro local, de vacas aneloradas não prenhes e transportados ao laboratório em bolsas plásticas à temperatura ambiente em solução salina (NaCl a 0,9%) entre 50-60 minutos. Nesse local os ovários foram lavados em álcool a 70% por 10 segundos e em seguida em solução salina. Primeiramente, foram lavados em solução salina e, em seguida, expostos a combinações de crioprotetores (VAJTA et al, 1998). Os folículos foram transferidos, imediatamente após sua retirada do nitrogênio, para uma solução de sacarose a 0,25 M, onde permaneceram por 2 minutos e, em seguida, transferidos para uma solução de sacarose a 0,15 M por mais 2 minutos e depois depositados em solução salina. Para o teste de toxicidade, os folículos foram expostos aos mesmos crioprotetores e períodos de tempo do grupo criopreservado, porém sem vitrificação. O liquido contendo os folículos foi centrifugado por 2 minutos a 1000 rpm, para a remoção dos crioprotetores - esse procedimento foi repetido duas vezes. Foram realizadas cinco repetições para cada tratamento (um ovário por repetição)
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