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Hum. Reprod. Advance Access published online on January 29, 2004
Human Reproduction, doi:10.1093/humrep/deh135
© 2004 by European Society of Human Reproduction and Embryology
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Received May 6, 2003
Revised August 14, 2003
Accepted December 4, 2003

Article

Derivation of a diploid human embryonic stem cell line from a mononuclear zygote

Edith Suss-Toby 1, S. Gerecht-Nir 2, M. Amit 3, D. Manor 1, and J. Itskovitz-Eldor 3*

1 Department of Obstetrics and Gynecology, Rambam Medical Center, POB 9602, Haifa 31096, Haifa, Israel
2 Department of Obstetrics and Gynecology, Rambam Medical Center, POB 9602, Haifa 31096, Haifa, Israel; Biotechnology-Interdisciplinary Unit, Technion-Israel Institute of Technology, Haifa, Israel
3 Department of Obstetrics and Gynecology, Rambam Medical Center, POB 9602, Haifa 31096, Haifa, Israel; Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel

* To whom correspondence should be addressed. E-mail: Itskovitz{at}rambam.health.gov.il.


  Abstract

BACKGROUND: IVF occasionally produces aneuploid zygotes with one or three pronuclei (PN). Routinely, these zygotes are discarded. The aim of this work was to establish human embryonic stem cell (hESC) lines from blastocysts resulting from abnormal fertilization. METHODS: Abnormally fertilized zygotes were cultured to the blastocyst stage and, following zona pellucida digestion, zona-free blastocysts were placed on a mouse feeder layer. Culture of hESCs was carried out as described earlier. RESULTS: Six out of the nine developing blastocysts attached to the feeder layer. One hESC line, originating from a mononuclear zygote following ICSI, was successfully derived. This line displayed typical phenotype and embryonic surface markers, and exhibited the potential to develop into all three embryonic germ layers both in vitro (by embryoid body formation) and in vivo (teratoma generation). Genetic examination revealed normal diploid karyotype and heterozygotic appearance for metachromatic leukodystrophy (MLD). CONCLUSION: This method, which requires neither immuno nor mechanical removal of the trophectoderm, may facilitate the derivation of hESC lines in general, and those from abnormal embryos in particular. Furthermore, it is shown that aneuploid zygotes can be used as a source for normal hESC derivation and hold the potential to generate aneuploid hESC lines for research purposes.

Key words: Key words: aneuploid zygote/derivation/differentiation/embryonic stem cells


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