The effects of permeating cryoprotectants on intracellular free-calcium concentrations and developmental potential of in vitro-matured feline oocytes
Jason R. Herrick A D , Chunmin Wang B C and Zoltan Machaty B
+ Author Affiliations
- Author Affiliations
A National Foundation for Fertility Research, 10290 RidgeGate Cr, Lone Tree, CO 80124, USA.
B Department of Animal Sciences, Purdue University, Lilly Hall, 915 West State St, West Lafayette, IN 47907, USA.
C Current Address: Vivere Health-Houston Surgery Center and IVF Laboratory, 2500 Fondren Rd, Suite 350, Houston, TX 77063, USA.
D Corresponding author. Email: jherrick@fertilityresearch.org
Reproduction, Fertility and Development 28(5) 599-607 https://doi.org/10.1071/RD14233
Submitted: 3 July 2014 Accepted: 26 August 2014 Published: 11 September 2014
Abstract
Embryos produced from vitrified feline oocytes have resulted in pregnancies, but the efficiency of oocyte vitrification in cats is still low. Our objectives were to evaluate the effects of exposing feline oocytes to ethylene glycol (EG), propanediol (PrOH) and dimethyl sulfoxide (DMSO) on changes in intracellular free-calcium concentrations ([Ca2+]i), the time needed for enzymatic digestion of the zona pellucida (ZP), the incidence of parthenogenetic activation and degeneration and embryonic development following in vitro fertilisation (IVF). All of the chemicals tested altered [Ca2+]i, but changes in [Ca2+]i, resistance of the ZP to enzymatic digestion and the incidence of parthenogenetic activation (<5% for all treatments) were not affected (P > 0.05) by extracellular Ca2+. Exposure to EG (>44.1%) and DMSO (19.7%) increased (P < 0.05) oocyte degeneration compared with control oocytes and oocytes exposed to PrOH (≤2.5%). Following exposure to a combination of PrOH and DMSO (10% v/v each), blastocyst development (per cleaved embryo; 52.1%) was similar (P > 0.05) to control oocytes (64.4%). When oocytes were vitrified with PrOH and DMSO, 28.3% of surviving (intact plasma membrane) oocytes cleaved following IVF, but no blastocyst developed. When a non-permeating cryoprotectant (galactose, 0.25 M) was added to the vitrification medium, 47.7% of surviving oocytes cleaved and 14.3% developed to the blastocyst stage.
Additional keywords: cat, DMSO, intracellular calcium, propanediol, vitrification.
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