Effects of Donor Cell Passage, Size and Type on Development of Porcine Embryos Derived from Somatic Cell Nuclear Transfer

Y H Zhang,E S Kim,P Q Cong,S H Lee,E S Song,C S Park,Y J Yi,J W Lee

doi:10.5713/ajas.2009.80344

Abstract

The aim of this study was to investigate the effects of donor cell passage, size and type on the development of nuclear transfer embryos. Porcine cumulus cells, fetal fibroblasts and oviductal epithelial cells from 1-2, 3-6 and 7-10 passages were used for the nuclear transfer. In the oocytes with the cumulus donor cells, fusion and cleavage rates of oocytes and cell numbers per blastocyst among the three different passage groups did not show any differences, but the rates of blastocyst formation from 1-2 and 3-6 passage groups were higher than those from 7-10 passage group. The rates of fusion, cleavage and blastocyst formation, and the cell numbers per blastocyst were higher in the embryos with the sizes of cumulus donor cells compared to the >20 cumulus donor cell. In the oocytes with the fetal fibroblast donor cells, the rate of blastocyst formation from the 3-6 passage group was higher than from 1-2 and 7-10 passage groups. The embryos with the size of 20 fetal fibroblast donor cell showed higher rate of blastocyst formation compared to those with 20 donor cells. In the oocytes with the oviductal epithelial cells, the rates of blastocyst formation from 1-2 and 3-6 passage groups were higher compared to those from 7-10 passage group. The embryos with the sizes of oviductal epithelial donor cells had a higher rate of blastocyst formation compared to those with >20 donor cell. Fusion and cleavage rates of oocytes, and cell numbers per blastocyst among the three different donor cell types from the 3-6 passage did not show any differences. However, the rate of blastocyst formation of somatic cell nuclear transfer (SCNT) embryos with the fetal fibroblast donor cell was higher than that of blastocyst formation of SCNT embryos with the cumulus and oviductal epithelial donor cells.

Highlights

The birth of cloned piglets by somatic cell nuclear transfer (SCNT) was reported in 2000 (Betthauser et al, 2000; Onishi et al, 2000; Polejaeva et al, 2000)
Effects of donor cell passage and size on development of embryos derived from somatic cell nuclear transfer
Wells et al (1999) reported that the number of donor cell passages did not show any differences on the development of somatic cell nuclear transfer (SCNT) embryos

Summary

Introduction

The birth of cloned piglets by somatic cell nuclear transfer (SCNT) was reported in 2000 (Betthauser et al, 2000; Onishi et al, 2000; Polejaeva et al, 2000). In the pig, the viability of SCNT embryos is poor, with an extremely low rate of cloned piglet production. Long-term cultured somatic cells undergo cellular senescence and have numerous mutations or allelic loss of genes accumulated through many rounds of cell divisions, which are known to cause improper genetic reprogramming after SCNT and subsequent abnormal development of the embryos (Aladjem et al, 1998; Cibelli et al, 1998; Kühholzer and Prather, 2000). According to the experiment results of Tao et al (1999), it was showed that donor cell size could affect the reprogramming of nuclear transfer embryos. The percentage of chromosome condensation and nuclear formation were higher in reconstructed embryos derived from small donor cells (15 μm) compared to large donor cells (20 μm), respectively

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