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Effect of elevated Ca 2+ concentration in fusion/activation medium on the fusion and development of porcine fetal fibroblast nuclear transfer embryos
Author(s) -
Cheong HeeTae,
Park KwangWook,
Im GiSun,
Lai Liangxue,
Sun QingYuan,
Day Billy N.,
Prather Randall S.
Publication year - 2002
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.10110
Subject(s) - blastocyst , andrology , embryo , fibroblast , electrofusion , perivitelline space , cell fusion , biology , oocyte activation , fetus , fusion , somatic cell nuclear transfer , embryo culture , in vitro , embryogenesis , oocyte , microbiology and biotechnology , cell , biochemistry , zona pellucida , materials science , pregnancy , medicine , genetics , linguistics , philosophy , metallurgy
The present study examined the effect of elevated Ca 2+ concentration in fusion/activation medium on the fusion and development of fetal fibroblast nuclear transfer (NT) porcine embryos. Frozen‐thawed and serum starved fetal fibroblasts were transferred into the perivitelline space of enucleated oocytes. Cell fusion and activation were induced simultaneously with electric pulses in 0.3 M mannitol‐based medium containing 0.1 or 1.0 mM CaCl 2 . Some fused embryos were further activated 1 hr after the fusion treatment by exposure to an electric pulse. The NT embryos were cultured in vitro for 6 days. Fusion and blastocyst formation rates were significantly ( P < 0.05) increased by increasing the Ca 2+ concentration from 0.1 mM (67.1 and 6.3%) to 1.0 mM (84.7 and 15.8%). However, no difference in the number of cells in blastocysts was observed between the two groups. A higher percentage of blastocyst was also observed when control oocytes were parthenogenetically activated in the presence of elevated Ca 2+ (19.3% vs. 32.4%, P < 0.05). When the reconstituted oocytes were fused in the medium containing 1.0 mM CaCl 2 , increasing the number of pulses from 2 to 3 or an additional activation treatment did not enhance the blastocyst formation rate or cell number in blastocysts. These results demonstrate that increasing the Ca 2+ concentration in the fusion/activation medium can enhance the fusion and blastocyst formation rates of fetal fibroblast NT porcine embryos without an additional activation treatment. Mol. Reprod. Dev. 61:488–492, 2002. © 2002 Wiley‐Liss, Inc.