M.R. Islam
Animal Division, National Institute of Biotechnology (NIB), Atomic Energy Research Establishment (AERE), EPZ, Savar, Dhaka-1349, Bangladesh
M.A.M.Y. Khandoker
Reproductive Biotechnology Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh
S. Afroz
Reproductive Biotechnology Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh
M.G.M. Rahman
Reproductive Biotechnology Laboratory, Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh
R.I. Khan
Department of Animal Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh
Goat ovary, Follicles, Cumulus-oocyte-complexes (COCs), In vitro production (IVP
Reproductive Biotechnology Laboratory, under the Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh, Bangladesh.
Animal Health and Management
The experiment was conducted at the Reproductive Biotechnology Laboratory, from Jan. 2004 to May 2005 under the Department of Animal Breeding and Genetics, Bangladesh Agricultural University, Mymensingh, Bangladesh.
Collection and processing of ovaries
Ovaries were collected from local slaughterhouse with their reproductive history being unknown. The ovaries were then recorded as right, left and the presence or absence of corpus luteum (CL) was also recorded. They were then kept in collection vial containing 0.9% physiological saline in a thermo flask at 25 °C to 30 °C and transported to the laboratory within 4 to 5 h of slaughter. The ovaries were then transferred to sterilized petridishes and rinsed thoroughly by physiological saline at 25 °C before further processing.
Measurement of weight, length and width
After trimming individually right, left, CL-present and -absent ovaries were weighed and the weight in gram was recorded in tabular form. The length and width in cm of the right, left, CL-present and -absent group ovaries were measured with the help of a measuring scale.
COCs aspiration and grading
The numbers of visible follicles on the surface of different category of ovaries were counted and recorded. The ovaries were washed 2 to 3 times in saline solution at 30 °C. Ten milliliters syringe was loaded with PBS (1~1.5 ml), and the needle (19 G) was put in the ovary parenchyma near the vesicular follicles of more than 2 mm diameter and all follicles were aspirated near the point. After aspirating the follicles from one ovary, the aspirated follicular materials were transferred slowly into a 90-mm petridish, avoiding damage of the cumulus cells and the COCs were searched and graded under microscope at low magnification. The COCs were classified according to the slight modification of the method of Khandoker et al.(2001) into 2 grades, normal: oocyte completely surrounded by cumulus cells; abnormal: oocyte partially surrounded by cumulus cells or completely denuded. The numbers of different grades of COCs in each category were recorded. In the meantime another petridish of Dulbeccos phosphate buffered saline (D-PBS) was prepared for pooling COCs and the COCs were picked up with an appropriate glass micropipette.
The tip diameter of the pipette was checked under the microscope to ensure COCs, which could be easily aspirated without damaging the cumulus cells. Basically the glass micropipettes were prepared slowly stretching the tip of Pasteur pipette above burners flame. Then the picked up COCs were washed 2 to 3 times into D-PBS.
All values were expressed as mean±SE. Statistical significance of differences between different parameters was evaluated by using Students unpaired t-test. The statistical analysis was done by SPSS (Statistical Package for Social Sciences, SPSS Inc., 1999; Microsoft Corporation, 1998) windows package and MINITAB.
J Zhejiang Univ Sci B. 2007 Jul; 8(7): 465–469
Journal