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Micromanipulation and cloning studies on buffalo oocytes and embryos using nucleus transfer
S K Singla1, R S Manik, M L Madan
1Embryo Biotechnology Centre, National Dairy Research Institute, Karnal, India.
Indian Journal of Experimental Biology
|May 6, 1998
Summary
This study demonstrates a successful new method for cloning buffalo embryos using nucleus transfer. The optimized enucleation and electrofusion techniques significantly improved cloned embryo development viability.
Area of Science:
- * Reproductive Biology
- * Animal Biotechnology
- * Embryology
Background:
- * Cloning buffaloes via somatic cell nuclear transfer (SCNT) is crucial for livestock improvement.
- * Efficient enucleation and electrofusion are key challenges in buffalo SCNT.
Purpose of the Study:
- * To develop and test a novel enucleation technique for buffalo oocytes.
- * To optimize electrofusion parameters for demi-oocyte-blastomere complexes.
- * To assess the viability of cloned buffalo embryos produced through this method.
Main Methods:
- * A new oocyte enucleation method involved zona cutting and pushing out half the cytoplasm with the nucleus.
- * Fluorescence screening confirmed successful enucleation (88.2%).
- * Optimized maturation time (37-42 hr), electrical pulses (3 pulses, 20V), and fetal bovine serum (20% FBS) for electrofusion and cleavage.
Main Results:
- * Maturation time significantly impacted fusion (37-42 hr better than 32 hr) and cleavage rates.
- * 3 electrical pulses and 20V resulted in higher fusion (54.1%) and cleavage (26.1%) rates.
- * 20% FBS supported optimal fusion (53.8%) and cleavage (23.2%) rates.
Conclusions:
- * The novel enucleation technique is viable for buffalo oocytes.
- * Optimized electrofusion parameters enhance cloned buffalo embryo development.
- * This study successfully produced cloned buffalo embryos, validating the technology.