Development Rates of Male Bovine Nuclear Transfer Embryos Derived from Adult and Fetal Cells1

Abstract

This study compared the nuclear transfer (NT) embryo development rates of adult and fetal cells within the same genotype. The adult fibroblast cells were obtained from a 21-yr-old Brahman bull. The fetal cells were derived from a Day 40 NT fetus previously cloned using cells from the Brahman bull. Overall, similar numbers of blastocysts developed from both adult (53 of 190; 28%) and fetal (39 of 140; 28%) donor cells. Improved blastocyst development rates were observed when fetal cells were serum-starved (serum-fed 12% vs. serum-starved 43%; P < 0.01) whereas there was no similar benefit when adult cells were serum-starved (both serum-fed and serum-starved 28%). Day 30 pregnancy rates were similar for blastocysts derived from adult (6 of 26; 23%) or fetal (5 of 32; 16%) cells. Day 90 pregnancy rates were 3 of 26 for adult and 0 of 32 for the fetal cell lines. One viable bull calf derived from a 21-yr-old serum-starved adult skin fibroblast was born in August 1999. In summary, somatic NT embryo development rates were similar whether adult or fetal cells, from the same genotype, were used as donor cells. Serum starvation of these adult donor cells did not improve development rates of NT embryos to blastocyst, but when fetal cells were serum-starved, there was a significant increase in development to blastocyst.

Development of swamp buffalo (Bubalus bubalis) embryos after parthenogenetic activation and nuclear transfer using serum fed or starved fetal fibroblasts

“…Our results were in agreement with the previous report in cattle that the nuclei of serum starved fibroblasts support the development of reconstructed embryos to the blastocyst stage significantly better than those of nonstarved fibroblasts (39% vs. 20%) [38]. Serum starvation of fetal donor cells before NT was also found to significantly improve NT blastocyst rates when compared with serum fed cells (43% vs. 12%) [39].…”

Comparative analysis of various donor cell types for somatic cell nuclear transfer and its association with apoptosis and senescence

“…This was in agreement with studies showing that the age, size, gender and type of donor cell may affect the efficiency of SCNT (4,(14)(15)(16). Moreover, culturing donor cells for long time periods resulted in decreased rates of in vitro embryos and to term development in cattle (17). Altogether, these studies indicate that nuclear donor cell preparation is a crucial step affecting the developmental potential of reconstructed embryos and the successful production of transgenic animals by SCNT.…”

Cell cycle analysis and interspecies nuclear transfer of in vitro cultured skin fibroblasts of the Siberian tiger (Panthera tigris Altaica)

“…Most of these efforts are focused on donor cells. These include: (a) synchrony of the cell cycle stage of donor cells (Kasinathan et al, 2001;Gibbons et al, 2002;Wells et al, 2003), as well as synchrony between donor cells and recipient oocytes (Campbell et al, 1994;Du et al, 2002); (b) using somatic cells from donors of various ages (Renard, 1999;Hill et al, 2000;Tian et al, 2000;Kasinathan et al, 2001;Xue et al, 2002), tissue origins (Galli et al, 1999;Shiga et al, 1999;Kato et al, 2000;Ogura et al, 2000;Du et al, 2002;Hochedlinger and Jaenisch, 2002;Miyashita et al, 2002), passages (Shiga et al, 1999;Arat et al, 2001;Liu et al, 2001) and culture conditions (Zakhartchenko et al, 1999b); (c) transfer of stem cells with low levels of epigenetic marks (Kato et al, 1999a;Wakayama et al, 1999;Amano et al, 2001;Eggan et al, 2001;Humpherys et al, 2001;Zhou et al, 2001); and (d) modifying epigenetic marks of donor cells with drugs (Jones et al, 2001;Zhou et al, 2002;Enright et al, 2003). Wilmut et al (2002) reported that a number of cellular factors influence the outcome of cloned offspring.…”