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-Meja, V.; Maleno, I.; Garca-Prez, J.L.; Menendez, P.
Source: Human Reproduction, Volume 24, Number 8, 29 August 2009 , pp. 1844-1851(8)
Publisher: Oxford University Press
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.
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 M 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.
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.
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.
Document Type: Research Article
Publication date: 2009-08-29
- Human Reproduction features full-length, peer-reviewed papers reporting original research, clinical case histories, as well as opinions and debates on topical issues. Papers published cover the scientific and medical aspects of reproductive physiology and pathology, endocrinology, andrology, gonad function, gametogenesis, fertilization, embryo development, implantation, pregnancy, genetics, genetic diagnosis, oncology, infectious disease, surgery, contraception, infertility treatment, psychology, ethics and social issues. The highest scientific and editorial standard is maintained throughout the journal along with a rapid rate of publication.
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- In this Subject: Anatomy & Physiology , Obstetrics & Gynecology
- By this author: Cortes, J.L. ; Sanchez, L. ; Ligero, G. ; Gutierrez-Aranda, I. ; Catalina, P. ; Elosua, C. ; Leone, P.E. ; Montes, R. ; Bueno, C. ; Ramos-Meja, V. ; Maleno, I. ; Garca-Prez, J.L. ; Menendez, P.