Derivation and characterisation of hESC lines from supernumerary embryos, experience from Odense, Denmark.
The derivation and characterisation of human embryonic stem cells provides a source of pluripotent stem cells with potential for clinical applications. Utilising locally sourced embryos from two IVF clinics, we derived and characterised five new cell lines for use in a non-clinical setting. Analysis of clinical data showed that the majority of embryos (94.5%) failed to reach the blastocyst stage of development and of all embryos, regardless of developmental status, 248 embryos were needed to create one stem cell line. From the number of embryos (69) which developed to the blastocyst stage 8.7% developed into cell lines. Using outgrowth of the whole blastocyst, we derived five new, unreported cell lines in Odense, Denmark between 2005 and 2006. Characterisation was carried out using RT-PCR, staining, karyotyping, EB formation and teratoma formation. The KMEB hESC lines will, in the future, be made available through the UK Stem Cell Bank (http://www.ukstemcellbank.org.uk/).
Harkness
Linda
L
Molecular Endocrinology laboratory (KMEB), Medical Biotechnology Centre, University of South Denmark, Winslowparken 25, 1 sal, 5000 Odense, Denmark. lharkness@health.sdu.dk
Rasmussen
Iben Anne
IA
Erb
Karin
K
Kassem
Moustapha
M
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D003114:Colony-Forming Units Assay; D003718:Denmark; D033041:Embryo Research; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D007621:Karyotyping; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013724:Teratoma / Q000473:pathology