Thesis Abstract and Dissertation Abstracts
Use of Simple Media For Supporting in Vitro Development of Buffalo Embryos
Abstract Category: Science
Course / Degree: M.V.Sc
Institution / University: National Dairy Research Institute, Karnal, India
Published in: 2005
In vitro production (IVP) of embryos, through a combination of the techniques of in vitro maturation (IVM), fertilization (IVF) and culture (IVC) of oocytes is a very important emerging reproductive technology in buffalo. However, complicated culture systems and very low blastocyst yields have limited its applicability.
The present study was, therefore, undertaken to i) compare the development of buffalo embryos in simple and complex culture media and ii) determine the effects of serum supplementation on the development of buffalo embryos.
Immature oocytes obtained from slaughterhouse buffalo ovaries were graded on the basis of their morphology. The usable quality oocytes were placed in 80-ìl droplets (15-20 oocytes/ droplet) of the IVM medium (TCM-199+10% FBS+5µg/ml porcine FSH+0.81mM sodium pyruvate+5% buffalo follicular fluid), covered with sterile paraffin oil in a 35 mm Petri dish and cultured for 24 h in a CO2 incubator (5% CO2 in air) at 38.5°C for carrying out IVM. Oocytes which exhibited cumulus expansion of degrees 1 and 2 were then subjected to IVF using standard protocols. After 18 h of sperm-oocyte incubation, the presumptive zygotes were divided randomly into the following 5 groups i) Controls; the zygotes were transferred to the control medium (TCM-199+10% FBS+co-culture with BOEC), which was changed at 18 and 48 hours post insemination (hpi) and then every 48 h, ii) CR; the zygotes were transferred to modified Charles-Rosenkrans-2 medium containing amino acids (mCR2aa) at 18 hpi, and the medium was changed at 48 hpi and then every 48 h, iii) CR+FBS; the zygotes were transferred to mCR2aa at 18 hpi and then to mCR2aa+10% FBS at 48 hpi. The medium was then changed every 48 h, iv) SOF; the zygotes were transferred to modified synthetic oviductal fluid containing amino acids (mSOFaa) at 18 hpi, and the medium was changed at 48 hpi and then every 48 h, and v) SOF+FBS; the zygotes were transferred to mSOFaa at 18 hpi and then to mSOFaa+10% FBS at 48 hpi. The medium was then changed every 48 h. The embryos were examined on Days 4, 6, 8 and 9 post insemination for evaluating their stage of development. The proportion of cleaved embryos that developed to the morula and blastocyst stages on Days 8 and 9 post insemination were compared among different groups by Chi square analysis.
Out of 697 oocytes subjected to IVM, 574 were fertilized, and 286 out of these were found to have cleaved at 48 hpi, with a cleavage rate of 50.5%. A comparison of the effects of the five media revealed that although there was no difference in the yields of Day 8 morulae and blastocysts, and Day 9 morulae, the cumulative figure of morulae + blastocysts at Day 8 post insemination was significantly higher.
Thesis Keywords/Search Tags:
Embryo, Simple Media
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Submission Details: Thesis Abstract submitted by Dharmendra Kumar from India on 06-Jan-2006 13:40.
Abstract has been viewed 2696 times (since 7 Mar 2010).
Dharmendra Kumar Contact Details: Email: vetdmkl@yahoo.co.in
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