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Hum. Reprod. Advance Access originally published online on November 7, 2007
Human Reproduction 2008 23(1):46-53; doi:10.1093/humrep/dem351
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© The Author 2007. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Laser-assisted derivation of human embryonic stem cell lines from IVF embryos after preimplantation genetic diagnosis

T. Turetsky1,{dagger}, E. Aizenman2,{dagger}, Y. Gil1, N. Weinberg3, Y. Shufaro1,2, A. Revel2, N. Laufer2, A. Simon2, D. Abeliovich3 and B.E. Reubinoff1,2,4

1 The Hadassah Human Embryonic Stem Cell Research Center, The Goldyne Savad Institute of Gene Therapy, Hadassah Hebrew University Medical Center, Ein Kerem, PO BOX 12000, Jerusalem 91120, Israel 2 Department of Obstetrics and Gynecology, Hadassah Hebrew University Medical Center, Ein Kerem, Jerusalem, Israel 3 Department of Human Genetics, Hadassah Hebrew University Medical Center, Ein Kerem, Jerusalem, Israel

4 Correspondence address. Tel: +972-2-6777495; Fax: +972-2-6430982; E-mail benjamin.reubinoff{at}ekmd.huji.ac.il

BACKGROUND: Human embryonic stem cells (hESCs) suitable for future transplantation therapy should preferably be developed in an animal-free system. Our objective was to develop a laser-based system for the isolation of the inner cell mass (ICM) that can develop into hESC lines, thereby circumventing immunosurgery that utilizes animal products.

METHODS: Hatching was assisted by micromanipulation techniques through a laser-drilled orifice in the zona pellucida of 13 abnormal preimplantation genetic diagnosed blastocysts. ICMs were dissected from the trophectoderm by a laser beam and plated on feeders to derive hESC lines.

RESULTS: eight ICMs were isolated from nine hatched blastocysts and gave rise to three hESC lines affected by myotonic dystrophy type 1, hemophilia A and a carrier of cystic fibrosis 405 + 1G > A mutation. Five blastocysts that collapsed during assisted hatching or ICM dissection were plated whole, giving rise to an additional line affected by fragile X. All cell lines expressed markers of pluripotent stem cells and differentiated in vitro and in vivo into the three germ layers.

CONCLUSIONS: These hESC lines can serve as an important model of the genetic disorders that they carry. Laser-assisted isolation of the ICMs may be applied for the derivation of new hESC lines in a xeno-free system for future clinical applications.

Key words: laser/preimplantation genetic diagnosis/human embryonic stem cells/derivation/inner cell mass

{dagger} T.T. and E.A. made equal contributions to this manuscript.

Submitted on July 9, 2007; resubmitted on September 20, 2007; accepted on October 8, 2007.


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