Effect of Donor Cell Types on Developmental Potential of Cattle (Bos taurus) and Swamp Buffalo (Bubalus bubalis) Cloned Embryos

Abstract

Abstract. This study investigated the effect of donor cell types on the developmental potential and quality of cloned swamp buffalo embryos in comparison with cloned cattle embryos. Fetal fibroblasts (FFs), ear fibroblasts (EFs), granulosa cells (GCs) and cumulus cells (CCs) were used as the donor cells in both buffalo and cattle. The cloned cattle or buffalo embryos were produced by fusion of the individual donor cells with enucleated cattle or buffalo oocytes, respectively. The reconstructed (cloned) embryos and in vitro matured oocytes without enucleation were parthenogenetically activated (PA) and cultured for 7 days. Their developmental ability to the blastocyst stage was evaluated. The total number of trophectoderm (TE) and inner cell mass (ICM) cells and the ICM ratio in each blastocyst was determined by differential staining as an indicator of embryo quality. The fusion rate of CCs with enucleated oocytes was significantly lower than for those of other donor cell types both in cattle and buffalo. The rates of cleavage and development to the 8-cell, morula and blastocyst stages of cloned embryos derived from all donor cell types did not significantly differ within the same species. However, the cleavage rate of cloned cattle embryos derived from FFs was significantly higher than those of cattle PA and cloned buffalo embryos. The blastocyst rates of cloned cattle embryos, except for the ones derived from CCs, were significantly higher than those of cloned buffalo embryos. In buffalo, only cloned embryos derived from CCs showed a significantly higher blastocyst rate than that of PA embryos. In contrast, all the cloned cattle embryos showed significantly higher blastocyst rates than that of PA embryos. There was no difference in ICM ratio among any of the blastocysts derived from any of the donor cell types and PA embryos in both species. FFs, EFs, GCs and CCs had similar potentials to support development of cloned cattle and buffalo embryos to the blastocyst stage with the same quality.

Effects of Trichostatin A on <i>In Vitro</i> Development and DNA Methylation Level of the Satellite I Region of Swamp Buffalo (<i>Bubalus bubalis</i>) Cloned Embryos

“…Donor cell and oocyte preparation, the somatic cell nuclear transfer procedure and in vitro embryo culture were performed as describe previously [1]. Briefly, buffalo oocytes were obtained from local abattoir-derived ovaries and cultured in in vitro maturation (IVM; TCM199 supplemented 10% FBS, 50 IU/ml hCG (Intervet, Boxmeer, Netherlands), 0.02 AU/ml FSH (Antrin ® , Kyoritsu Seiyaku, Tokyo, Japan) and 1 µg/ml 17β-estradiol) medium at 38.5 C under a humidified atmosphere of 5% CO 2 in air for 23 h. Then, cumulus cells were removed from cumulus oocytes complexes (COC) by repeat pipetting in 0.2% hyaluronidase.…”

“…From our previous report, the developmental potential to the blastocyst stage of swamp buffalo cloned embryos was lower than that of bovine cloned embryos [1]. Increasing the blastocyst rate in swamp buffalo cloned embryos enhances the possibility to obtain viable clones.…”

A comparative study on efficiency of adult fibroblasts and amniotic fluid-derived stem cells as donor cells for production of hand-made cloned buffalo (Bubalus bubalis) embryos

“…The blastocyst formation of embryos derived from cumulus cells was higher than those of embryos derived from fetal fibroblast or adult fibroblast (Shah et al 2009). The donor cell types, adult fibroblast, fetal fibroblast or cumulus cells had similar ability to support cleavage and embryo development (Srirattana et al 2010). The adult fibroblast derived from ear pinna and NT-ES cell-like cells derived from cloned blastocysts generated using this adult fibroblast as donor cells, when used for NT, gave almost similar cleavage and blastocyst rates (George et al 2011).…”

Handmade Cloned Buffalo (Bubalus bubalis) Embryos Produced from Somatic Cells Isolated from Milk and Ear Skin Differ in Their Developmental Competence, Epigenetic Status, and Gene Expression

“…Of the more than 200 distinct mammalian cell types known, less than 5% have been tested as nuclear donors, and all of them have been found to support development to blastocyst stage (Oback and Wells, 2002). In all the studies in which the cloning efficiency of different cell types was compared within a species, it was found to vary substantially in cattle (Kato et al, 2000), swamp buffalo (Srirattana et al, 2010), and pig (Zhang et al, 2009).…”