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Selected Publication:

Type of publication: Journal Article
Type of document: Full Paper

Year: 1999

Authors: Zakhartchenko, V; Alberio, R; Stojkovic, M; Prelle, K; Schernthaner, W; Stojkovic, P; Wenigerkind, H; Wanke, R; Düchler, M; Steinborn, R; Mueller, M; Brem, G; Wolf, E

Title: Adult cloning in cattle: Potential of nuclei from a permanent cell line and from primary cultures.

Source: Mol Reprod Dev 1999; 54(3): 264-272.

Authors Vetmeduni Vienna:

Brem Gottfried
Steinborn Ralf

Vetmed Research Units
Institute of Animal Breeding and Genetics

Nuclear transfer was used to evaluate the developmental potential of nuclei from a spontaneously immortalized bovine mammary gland epithelial cell line (MECL) and from primary cultures of mammary gland cells (PMGC) and ear skin fibroblasts (PESF) established from 3-year-old cows. Cell proliferation was investigated by incorporation and detection of 5-bromo-2"-deoxyuridine (BrdU). The proportion of cells in S-phase was significantly (P < 0.05) higher for MECL cells than for PMGC and PESF, both in the presence of serum (90% vs. 28% and 15%) and following serum starvation (27% vs. 6% and 3%). Nuclei from PESF supported the development of reconstructed embryos to the blastocyst stage significantly better than those of PMGC (60% vs. 26%; P < 0.05). Embryos reconstructed with cells from MECL failed to develop to blastocysts. After transfer of embryos derived from PMGC and PESF, respectively, 2/2 and 5/12 recipients were pregnant on day 42. On day 90, the corresponding pregnancy rates were 2/2 and 3/12. One live calf derived from a PMGC was born at day 287 of gestation. Another live PESF-derived calf was delivered by caesarean section at day 286 of gestation. Our study suggests that nuclei from primary cultures of adult cells can be successfully reprogrammed by nuclear transfer, whereas nuclei from a permanent cell line failed to support the development of nuclear transfer embryos.

Keywords Pubmed: Animals
Cell Culture Techniques/methods*
Cell Division
Cell Line
Cell Nucleus/genetics*
Cell Nucleus/metabolism
Cloning, Molecular/methods*
Culture Media, Serum-Free
Embryo Transfer/methods*
Nuclear Transfer Techniques*
Time Factors

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