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Nuclear transfer using a bovine mammary epithelial cell line (BMEC)
Author(s) -
AKAGI Satoshi,
TAKAHASHI Seiya,
OHKOSHI Katsuhiro,
TAKENOUCHI Takato,
SHIMIZU Manabu,
GESHI Masaya,
ADACHI Noritaka,
FUCHIMOTO Daiichiro,
IZAIKE Yoshiaki,
ASO Hisashi
Publication year - 2002
Publication title -
animal science journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.606
H-Index - 38
eISSN - 1740-0929
pISSN - 1344-3941
DOI - 10.1046/j.1344-3941.2002.00064.x
Subject(s) - blastocyst , embryo , oocyte , cell fusion , fusion , cell culture , chemistry , embryo transfer , cell , biology , microbiology and biotechnology , embryogenesis , biochemistry , genetics , linguistics , philosophy
The developmental potential of nuclei from a bovine mammary epithelial cell line (BMEC) in nuclear transfer was investigated. For nuclear transfer donors, BMEC cells (passage 15) were cultured for 4–5 days after seeding at cell densities of 1.0 × 10 5 cells/cm 2 (high‐density group) or 0.8 × 10 4 cells/cm 2 (low‐density group). First, the effective electric stimulation for fusion of enucleated oocytes with BMEC cells was examined. Fusion rates reached maximum with two DC pulses of 30 V/150 µm for 20 µs in the high‐density group and with two DC pulses of 25 V/150 µm for 10 µs in the low‐density group. The fusion rate (37.5%) in the high‐density group was significantly ( P < 0.005) lower than in the low‐density group (71.4%). Second, the in vitro developmental potential of nuclear transfer embryos derived from BMEC cells was examined. In the high‐density and low‐density groups, 18.8% and 24.1% of fused oocytes, respectively, developed to blastocyst stage. The results of this study indicate that nuclei from BMEC cells support the development of nuclear transfer embryos to the blastocyst stage and that the efficiency of oocyte–cell fusion is affected by the culture conditions of the donor BEMC cells before nuclear transfer.