Diploid, but not haploid, human embryonic stem cells can be derived from microsurgically repaired tripronuclear human zygotes

Abstract

(2013) Diploid, but not haploid, human embryonic stem cells can be derived from microsurgically repaired tripronuclear human zygotes, Cell Cycle, 12:2, 302-311,

Improved Efficiency of Microsurgical Enucleated Tripronuclear Zygotes Development and Embryonic Stem Cell Derivation by Supplementing Epidermal Growth Factor, Brain-Derived Neurotrophic Factor, and Insulin-Like Growth Factor-1

“…However, since all hESC lines have chromosomal abnormalities, the risks involved in this method need to be evaluated [33]. In our previous study, we also indicated that the diploid hESC lines can be derived from 3PN zygotes when one pronuclear is removed [8], and Jiang et al subsequently proved our results [9]; however, the lower efficiency limited the development of this method. Here, we have developed a new culture system to promote the application of 3PN zygotes.…”

“…Very often (5%-7% incidence rate), two sperms enter the oocyte, forming polyspermic zygotes (also named 3PN zygotes, because three pronuclear ones are observed in these zygotes). In a previous study, Fan et al demonstrated that the cytoplasm of 3PN zygotes can reprogram human somatic cells by nuclear transfer technology, indicating that the cytoplasm of 3PN zygotes is comparable to that of normal zygotes [8], and these results were further corroborated by Kawamura et al [24]. Therefore, the concern was whether diploid zygotes could be obtained after the removal of a redundant pronucleus.…”

“…ICM was isolated by the mechanical method as described in our previous study [8]. After 5-7 days, the human ICM formed a small colony, and the culture medium was changed every 2 days.…”

“…Kattera and Chen transferred repaired embryos into a 38-year-old woman, resulting in a normal, healthy baby boy [7]. Based on these findings, Fan et al derived the first hESC line using five blastocysts developed from 3PN zygotes repaired by microsurgery [8]. Subsequently Jiang et al obtained 2 hESC lines using 11 blastocysts generated by the microsurgical method [9].…”

“…In previous studies, the lower quality of polyspermic zygotes has been shown to result in lower blastocyst formation efficiency and low blastocyst quality. Therefore, improvements in embryo quality could contribute to an increase in hESC derivation [8].…”