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- Title
Mesenchymal stem cells facilitate the derivation of human embryonic stem cells from cryopreserved poor-quality embryos.
- Authors
Cortes, J.L.; Sanchez, L.; Ligero, G.; Gutierrez-Aranda, I.; Catalina, P.; Elosua, C.; Leone, P.E.; Montes, R.; Bueno, C.; Ramos-Mejía, V.; Maleno, I.; García-Pérez, J.L.; Menendez, P.
- Abstract
: BACKGROUND Human embryonic stem cells (hESCs) have opened up a new area of research in biomedicine. The efficiency of hESC derivation from frozen poor-quality embryos is low and normally achieved by plating embryos on mouse or human foreskin feeders (HFFs). We attempted to optimize embryo survival and hESC derivation. : METHODS Three conditions were tested on frozen poor-quality embryos: (i) embryo treatment with the Rho-associated kinase (ROCK) inhibitor, Y-27632; (ii) use of human mesenchymal stem cells (hMSCs) as feeders; and (iii) laser drilling (LD) for inner cell mass (ICM) isolation. Two hundred and nineteen thawed embryos were randomly treated with (n = 110) or without (n = 109) 10 µM Y-27632. Surviving embryos that developed to blastocyst stage (n = 50) were randomly co-cultured on HFFs (n = 21) or hMSCs (n = 29). ICM isolation was either by whole-blastocyst culture (WBC) or WBC plus LD. : RESULTS Embryo survival was 52% higher with Y-27632. hMSCs appeared to facilitate ICM outgrowth and hESC derivation: three hESC lines were derived on hMSCs (10.3% efficiency) whereas no hESC line was derived on HFFs. ROCK inhibition and ICM isolation method did not affect hESC efficiency. The lines derived on hMSCs (AND-1, -2, -3) were characterized and showed typical hESC morphology, euploidy, surface marker and transcription factor expression and multilineage developmental potential. The hESC lines have been stable for over 38 passages on hMSCs. : CONCLUSION Our data suggest that Y-27632 increases post-thaw embryo survival and that hMSCs may facilitate the efficiency of hESC derivation from frozen poor-quality embryos.
- Subjects
EMBRYONIC stem cells; MESENCHYME; EMBRYOS; CRYOPRESERVATION of organs, tissues, etc.; BLASTOCYST; CELL culture
- Publication
Human Reproduction, 2009, Vol 24, Issue 8, p1844
- ISSN
0268-1161
- Publication type
Article
- DOI
10.1093/humrep/dep107