In vitro fertilisation of in vivo matured porcine oocytes obtained from prepuberal gilts at different time intervals after hCG injection

Rátky, J. and Rath, D. and Brüssow, K.-P. (2003) In vitro fertilisation of in vivo matured porcine oocytes obtained from prepuberal gilts at different time intervals after hCG injection. Acta Veterinaria Hungarica, 51 (1). pp. 95-101. ISSN 0236-6290

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Abstract

The goal of the present study was to find out the best interval after hCG injection in PMSG primed prepuberal gilts for retrieval of in vivo matured oocytes for in vitro fertilisation (IVF). Altogether 66 gilts were superovulated with 1500IU PMSG and 500IU hCG 72h later. Ovum pick up was performed endoscopically 24, 28, 32 or 36h after hCG and a total of 869 cumulus-oocyte-complexes (COCs) were aspirated from 1400 follicles. COCs were tested for quality, and an aliquot was immediately fixed and stained to determine meiotic configuration. The remaining COCs were fertilised in vitro using frozen-thawed epididymal semen. Quality and developmental stage of embryos were tested after IVF, and the number of nuclei was counted. At 24 to 32h after hCG only few oocytes have entered the second meiotic cycle (18 to 25% vs. 58% at 36h, p<0.05). The overall cleavage rate was significantly influenced by insufficient maturation rate at the early collection times (14% at 24h vs. 49% at 36h). Additionally, when oocytes were collected 24 to 32h vs. 36h the cleavage rate based on mature oocytes was lower (26 vs. 62%, p<0.05). Once embryonic development has been initiated, the further in vitro development to blastocyst stages did not differ between groups. However, the number of cells was lower at collection times 24 to 32h as compared to 36h after hCG (12 to 15 cells vs. 22 cells, p<0.05). The results indicate that the time of COC collection affects the in vitro developmental competence up to the blastocyst stage and should not be performed earlier than 36h after hCG treatment.

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