Microtubular spindle dynamics and chromosome complements from somatic cell nuclei haploidization in mature mouse oocytes and developmental potential of the derived embryos

Shee Uan Chen, Chia Yi Chang, Chien Cheng Lu, Fon Jou Hsieh, Hong Nerng Ho, Yu Shih Yang

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background: The aim of this study was to investigate haploidization of somatic cell nuclei in non-enucleated mature oocytes regarding spindle formation, chromosomes and developmental potential. Methods: Mouse cumulus cells were injected into metaphase II oocytes. Some injected oocytes were examined for morphological changes of chromosomes and the spindle immediately, and at 30 min, 1 h or 2 h after the injections. The remaining oocytes were activated by Sr 2+ after various incubation periods and observed for formation of a second polar body and pseudo-polar body. Cytogenetic analysis was performed for some of the resulting zygotes. The progress to blastocysts in vitro and the possibility of conception in vivo were assessed. Results: Immediately after injection, the cumulus cell nucleus was still in interphase without spindle formation. The occurrence of premature chromosome condensation (PCC) and spindle formation increased as the incubation time increased. The percentages of activated oocytes increased with the incubation time after nuclear transfer, but the difference was not significant between 1 (58%) and 2 h (62%). The incidence of chromosomal aberrations was high for the derived embryos. Development in vitro was poor, and no procreation of pups occurred after transfer of the 324 embryos. Conclusions: The PCC and spindle formation induced by cumulus cell nuclei in mature oocytes was time dependent, as was the chance for successful activation. The chromosomal abnormalities from segregation errors presented one obvious cause, apart from the potential epigenetic defects, of developmental failure of the semi-cloned embryos.

Original languageEnglish
Pages (from-to)1181-1188
Number of pages8
JournalHuman Reproduction
Volume19
Issue number5
DOIs
Publication statusPublished - Jan 1 2004
Externally publishedYes

Fingerprint

Cell Nucleus
Oocytes
Embryonic Structures
Chromosomes
Cumulus Cells
Polar Bodies
Chromosome Aberrations
Injections
Zygote
Cytogenetic Analysis
Embryo Transfer
Interphase
Blastocyst
Metaphase
Epigenomics
Incidence

Keywords

  • Chromosomes
  • Haploidization
  • Microtubular spindle
  • Nuclear transfer

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynaecology

Cite this

Microtubular spindle dynamics and chromosome complements from somatic cell nuclei haploidization in mature mouse oocytes and developmental potential of the derived embryos. / Chen, Shee Uan; Chang, Chia Yi; Lu, Chien Cheng; Hsieh, Fon Jou; Ho, Hong Nerng; Yang, Yu Shih.

In: Human Reproduction, Vol. 19, No. 5, 01.01.2004, p. 1181-1188.

Research output: Contribution to journalArticle

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AB - Background: The aim of this study was to investigate haploidization of somatic cell nuclei in non-enucleated mature oocytes regarding spindle formation, chromosomes and developmental potential. Methods: Mouse cumulus cells were injected into metaphase II oocytes. Some injected oocytes were examined for morphological changes of chromosomes and the spindle immediately, and at 30 min, 1 h or 2 h after the injections. The remaining oocytes were activated by Sr 2+ after various incubation periods and observed for formation of a second polar body and pseudo-polar body. Cytogenetic analysis was performed for some of the resulting zygotes. The progress to blastocysts in vitro and the possibility of conception in vivo were assessed. Results: Immediately after injection, the cumulus cell nucleus was still in interphase without spindle formation. The occurrence of premature chromosome condensation (PCC) and spindle formation increased as the incubation time increased. The percentages of activated oocytes increased with the incubation time after nuclear transfer, but the difference was not significant between 1 (58%) and 2 h (62%). The incidence of chromosomal aberrations was high for the derived embryos. Development in vitro was poor, and no procreation of pups occurred after transfer of the 324 embryos. Conclusions: The PCC and spindle formation induced by cumulus cell nuclei in mature oocytes was time dependent, as was the chance for successful activation. The chromosomal abnormalities from segregation errors presented one obvious cause, apart from the potential epigenetic defects, of developmental failure of the semi-cloned embryos.

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