The viability of oocytes should be a important selection parameter for successful in vitro culture systems. This study evaluated the viability of canine oocytes before and during in vitro culture period.
Material & Methods
Bitch oocytes were collected from ovariohysterectomy; the ovaries were sliced and released oocytes were washed with PBS The oocytes were placed in 50µl drops of maturation media: (TCM 199 with 10% fetal calf serum, 20 mM Hepes and 10 UI/ mL of hCG) for 72 h at 39°C with 5% CO2 in air. Oocyte viability was assessed by fluorescein diacetate staining (FDA) under epifluorescence microscope before cultured and every 24 h during culture period. Green fluorescence indicated cell viability.
Before culture 399/450 (88,6%) showed fluorescence; during culture period the cell viability was: 62/67 (92.5%); 63/73 (86.3%) and 80 /113 (70.7%), at 24, 48 and 72 h respectively.
A high percentage of cell viability can be obtained from canine ovaries which are maintained during culture in vitro.
Supported by Grant 1030380 FONDECYT.