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KMID : 0900920030270010053
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Korean journal of Animal Reproduction
2003 Volume.27 No. 1 p.53 ~ p.59
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Sexing and Cell Cycle Induction Hanwoo Fetal Fibroblast Cells
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Kim Hyun-Joo
Kang Hyee-Sung
Choi Hwa-Sik
Lee Seung-Ho
Park Chang-Sik
Jin Dong-Il
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Abstract
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For somatic cell nuclear transfer in Hanwoo, fetal fibroblast cell lines were established from 35, 50, 70 and 90-day fetuses of Korean native cattle. The sex of these fetal fibroblast cells were analyzed by PCR using Y-specific primers and confirmed that two cell lines were female and the other two cell lines were male. Karyotyping of these cell lines indicates that the chromosome numbers of fetal fibroblast cells were not affected by passage number and more than 80% of fetal fibroblast cells have normal chromosome number. To evaluate Go stage in cell cycle of fetal fibroblast cells, Western blotting was performed to detect the expression level of PCNA which is known to be expressed in all cell cycle stages except G_{0} stage. Following serum starvation or confluent culture for 7 days, fetal fibroblast cells were effectively reached to G_{0} stage. The cell cycle was resumed after culture of these Go stage-fetal fibroblast cells with normal medium. These results indicates that fetal fibroblast cells originated from Hanwoo were successfully isolated and culture system and induction of cell cycle of these cells were established for somatic cell nuclear transfer in Hanwoo.woo.
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KEYWORD
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Hanwoo fetal fibroblast cells, PCR sexing, Karyotyping, PCNA expression
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