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

Year: 1999

Authors: Zakhartchenko, V; Durcova-Hills, G; Stojkovic, M; Schernthaner, W; Prelle, K; Steinborn, R; Müller, M; Brem, G; Wolf, E

Title: Effects of serum starvation and re-cloning on the efficiency of nuclear transfer using bovine fetal fibroblasts.

Source: J Reprod Fertil (115), 2 325-331.

Authors Vetmeduni Vienna:

Brem Gottfried
Müller Mathias
Steinborn Ralf

Vetmed Research Units
Institute of Animal Breeding and Genetics

The developmental potential of bovine fetal fibroblasts was evaluated using nuclear transfer. Fibroblasts from a 37-day-old fetus were fused to enucleated oocytes before activation. Nuclei of starved (cultured for 8 days in medium containing 0.5% serum) fibroblasts supported the development of reconstructed embryos to the blastocyst stage significantly better than those of non-starved fibroblasts (39% versus 20%; P < 0.05). When nuclear transfer morulae derived from starved or non-starved fibroblasts were used for re-cloning, the proportion of blastocysts (52 and 55%, respectively) obtained with these embryonic nuclei was significantly higher than it was with fibroblast nuclei used in the first round of nuclear transfer (P < 0.05 and P < 0.001, respectively). After transfer of blastocysts derived from non-starved and starved fibroblasts, respectively, 33% (1/3) and 78% (7/9) of recipients were pregnant on day 30 as assessed by ultrasonography. On day 90, the corresponding pregnancy rates were 33% (1/3) and 63% (5/8). Two live male twin calves, derived from non-starved fibroblasts, were delivered by Caesarean section at day 281 of gestation. This study demonstrates a positive effect of serum starvation on the efficiency of nuclear transfer using bovine fetal fibroblasts. The efficiency of nuclear transfer could be further increased by recloning.

Keywords Pubmed: Animals
Cells, Cultured
Chi-Square Distribution
Clone Cells*
Microscopy, Fluorescence
Nuclear Transfer Techniques*

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