Vitrification of mouse embryo-derived ICM cells: a tool for preserving embryonic stem cell potential?
OBJECTIVE
Vitrification technology presents new opportunities for preservation of embryo derived stem cells without first establishing a viable ESC line. This study tests the feasibility of cryopreserving ICM cells using vitrification.


METHODS
ICMs from mouse embryos were isolated and vitrified in HSV straws or on cryoloops. Upon warming, the vitrified ICMs were cultured and observed for attachment and morphology. Colonies were passaged every 3-6 days. ICMs and ICM-derived ESC colonies were tested for expression of stem cell specific markers.


RESULTS
ICMs vitrified on both the cryoloop and the HSV straw had high survival rates. ICM derived ESCs remained undifferentiated for several passages and demonstrated expression of typical stem cell markers; SSEA-1, Sox-2, Oct 4 and alkaline phosphatase.


CONCLUSIONS
This is the first report on successful vitrification of isolated ICMs and the subsequent derivation of ESC colonies. Vitrification of isolated ICMs is a novel approach for preservation of the "stem cell source" material.
Department of OB-GYN, Cleveland Clinic Foundation, Women's Health Institute, Beachwood, OH, USA. desain@ccf.org
Nina
N
Desai
Jing
J
Xu
Tamara
T
Tsulaia
Julia
J
Szeptycki-Lawson
Faten
F
AbdelHafez
James
J
Goldfarb
Tommaso
T
Falcone
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D053624:Blastocyst Inner Cell Mass / Q000166:cytology*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D015925:Cryopreservation / Q000379:methods; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005109:Extracellular Matrix; D016256:Lewis X Antigen / Q000032:analysis; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000032:analysis; D058989:Vitrification
D016256:Lewis X Antigen; D050814:Octamer Transcription Factor-3; D000469:Alkaline Phosphatase