Self-correction of chromosomally abnormal embryos in culture and implications for stem cell production

Munné, Santiago; Velilla, Esther; Colls, Pere; Bermudez, Mercedez Garcia; Vemuri, Mohan C.; Steuerwald, Nury; Garrisi, John; Cohen, Jacques

doi:10.1016/j.fertnstert.2005.06.025

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Volume 84, Issue 5 , Pages 1328-1334, November 2005

Self-correction of chromosomally abnormal embryos in culture and implications for stem cell production

Santiago Munné, Ph.D.
- Reprogenetics, LLC, West Orange, New Jersey, USA
- Reprint requests: Santiago Munné, Ph.D., Reprogenetics, LLC, 101 Old Short Hills Road, Suite 501, West Orange, New Jersey 07052 (FAX: 973-3226235).
Esther Velilla, M.Sc.
- Reprogenetics, LLC, West Orange, New Jersey, USA
Pere Colls, Ph.D.
- Reprogenetics, LLC, West Orange, New Jersey, USA
Mercedez Garcia Bermudez, Ph.D.
- Reprogenetics, LLC, West Orange, New Jersey, USA
Mohan C. Vemuri, Ph.D.
- Reprogenetics, LLC, West Orange, New Jersey, USA
Nury Steuerwald, Ph.D.
- A.R.T. Institute of New York and New Jersey, West Orange, New Jersey, USA
John Garrisi, Ph.D.
- Saint Barnabas Medical Center, Livingston, New Jersey
Jacques Cohen, Ph.D.
- Reprogenetics, LLC, West Orange, New Jersey, USA

Received 7 February 2005; received in revised form 30 June 2005; accepted 30 June 2005.

Objective

To ascertain whether embryos classified by preimplantation genetic diagnosis (PGD) for infertility as abnormal and then plated to obtain stem cells would self-correct partially or totally in culture, producing disomic stem cells.

Design

Prospective study to determine the chromosome status of embryos on day 3 and 6, as well as cultured cells derived from inner cell masses from the same embryos when cultured up to day 12.

Setting

Research laboratory.

Patient(s)

Patients undergoing PGD of aneuploidy.

Intervention(s)

Of 142 embryos classified by PGD for aneuploidy as abnormal, 50 were cultured to the blastocyst stage. At that stage a fraction of the embryos underwent trophectoderm biopsy to reconfirm the PGD diagnosis. After further co-culture with feeders up to day 12, 34 embryos attached to the feeder cells. Of those, 24 were analyzed by fluorescence in situ hybridization (FISH) and the rest for the expression of Oct-4, SSEA-3, SSEA-4, TRA1-60, and TRA1-80.

Main Outcome Measure(s)

Disomic cells obtained from trisomic embryos.

Result(s)

Analysis by FISH of day-12 cultures showed that 7 were totally normal, 6 were mostly abnormal, and 11 had experienced some chromosome normalization, having between 21% and 88% normal cells. Day-12 culture was positive for Oct-4 expression by reverse transcriptase polymerase chain reaction analysis and for SSEA-3, SSEA-4, TRA1-60, and TRA1-80 by immunocytochemistry.

Conclusion(s)

Chromosome self-normalization occurs in a significant proportion of chromosomally abnormal embryos, possibly because of the loss of a chromosome in trisomic cells after blastocyst stage. Thus chromosomally abnormal embryos are a potential source of disomic stem cells. Not all chromosomally abnormal embryos self-corrected. Abnormal stem cells that might be derived could be used as models to study the effect of chromosomal abnormalities on human development.

Key Words: Trisomy , preimplantation genetic diagnosis , mosaicism