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International Journal of Molecular Veterinary Research
2013, Vol.3, No.2, 4
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saline (0.9%) containing gentamicin (50 µg/mL). The
genitalia were washed with tap water thrice and the
ovaries were separated. The separated ovaries were
sorted as left and right ovaries and washed separately
with DPBS (pH 7.3) containing gentamicin (50 µg/mL).
2.1 Oocyte recovery
The oocytes were collected aseptically from the
ovaries by three methods:
(1) Aspiration: The visible follicles present on the
surface of the ovary were aspirated with 22 G needle
fixed to 5 mL disposable syringe containing 1~2 mL
of DPBS. The goat oocytes were aspirated from
individual ovaries after carefully removing the
extraneous tissue and were placed in petri dish
containing 1 mL of DPBS for grading.
(2) Dissection Technique: The ovaries were placed in
a sterile glass petri dish containing 2 mL of DPBS. All
the visible follicles were carefully subjected to blunt
dissection with the help of forceps and the remaining
ovarian tissue was removed after a brief rinsing. The
follicles were ruptured and the follicular fluid was
allowed to flow into the DPBS.
(3) Slicing: The ovaries were held firmly with the help
of forceps in a sterile glass petri dish containing 2 mL
of DPBS. The ovaries were sliced into possible thin
sections with a blade fixed to the artery forceps. The
oocytes containing DPBS media were transferred to
the petri dish and observed under Stereozoom
microscope to grade the oocytes.
The petri dish was observed under Stereozoom
microscope and the oocytes were transferred to a
searching dish containing DPBS for grading.
2.2 Grading of oocytes
Oocytes were classified on the basis of cumulus layer as
follows (Table 1; Figure 4) (Singh and Sharma, 1991).
Table 1 Classification grading of oocytes
No.
Number of cumulus layers present
Grade
1
More than three layers of cumulus cells
A
2
One to two layers of cumulus cells
B
3
Scattered cumulus complex
C
Figure 4 A, B, C and D grades of oocytes in non descriptive dose
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