Effects of donor cell type and genotype on the efficiency of mouse somatic cell cloning
Effects of donor cell type and genotype on the efficiency of mouse somatic cell cloning
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DOI：
10.1095/biolreprod.103.017731
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发表时间：
2003-10-01
影响因子：
3.6
通讯作者：
Ogura, A
中科院分区：
文献类型：
--
作者：
Inoue, K;Ogonuki, N;Ogura, A
虽然人们普遍认为供体细胞的细胞类型和基因型影响体细胞克隆的效率，但很少有系统的分析来验证这一假设。进行本研究以检查供体细胞类型、供体基因型或其组合是否增加小鼠克隆的效率。最初，我们评估了从具有六种不同基因型的卵丘或未成熟支持细胞克隆的胚胎的发育能力（即，2 X 6阶乘）。卵丘细胞的卵裂率显著高于支持细胞（P < 0.005，双因素方差分析），这可能是由于卵丘细胞在G 0/G1期的上级细胞周期同步性较好。胚胎移植后，细胞类型对出生率有显著影响，支持细胞的效果更好（P < 0.005）。细胞类型与基因型之间存在显著的交互作用（P < 0.05），表明克隆效率是由细胞类型和基因型共同决定的。使用（136 x 129）F1 Sertoli细胞获得的平均出生率最高（10.8 +/- 2.1%）。在第二系列实验中，我们检测了当与129基因型组合时，具有野生型基因型（JF 1）的克隆的发育能力是否得到改善。正常幼崽是从（129 x JF 1）F1和（JF 1 x 129）F1基因型的卵丘和未成熟支持细胞克隆的，而没有幼崽是从（136 x JF 1）F1基因型的细胞中出生的。本研究清楚地表明，体细胞克隆的效率，特别是胚胎移植后的胎儿存活率，可以通过选择适当的细胞类型和基因型的组合显着提高。
Although it is widely assumed that the cell type and genotype of the donor cell affect the efficiency of somatic cell cloning, little systematic analysis has been done to verify this assumption. The present study was undertaken to examine whether donor cell type, donor genotype, or a combination thereof increased the efficiency of mouse cloning. Initially we assessed the developmental ability of embryos that were cloned from cumulus or immature Sertoli cells with six different genotypes (i.e., 2 X 6 factorial). Significantly better cleavage rates were obtained with cumulus cells than with Sertoli cells (P < 0.005, two-way ANOVA), which probably was due to the superior cell-cycle synchrony of cumulus cells at G0/G1. After embryo transfer, there was a significant effect of cell type on the birth rate, with Sertoli cells giving the better result (P < 0.005). Furthermore, there was a significant interaction (P < 0.05) between the cell type and genotype, which indicates that cloning efficiency is determined by a combination of these two factors. The highest mean birth rate (10.8 +/- 2.1%) was obtained with (136 x 129) F1 Sertoli cells. in the second series of experiments, we examined whether the developmental ability of clones with the wild-type genotype (JF1) was improved when combined with the 129 genotype. Normal pups were cloned from cumulus and immature Sertoli cells of the (129 x JF1)F1 and (JF1 x 129)F1 genotypes, whereas no pups were born from cells with the (136 x JF1)F1 genotype. The present study clearly demonstrates that the efficiency of somatic cell cloning, and in particular fetal survival after embryo transfer, may be improved significantly by choosing the appropriate combinations of cell type and genotype.