Establishment of hESC Lines from the Inner Cell Mass of Blastocyst-Stage Embryos and Single Blastomeres of 4-Cell Stage Embryos

More than 600 human embryonic stem cell (hESC) lines have been reported today at the human European Embryonic Stem Cell Registry (http://www.hescreg.eu/ ). Despite these high numbers, there are currently no general protocols for derivation, culture, and characterization of hESC. Moreover, data on the culture of the embryo used for the derivation (medium, day of ICM isolation) are usually not available but can have an impact on the derivation rate. We present here the protocols for derivation, culture and characterization as we applied them for the 22 hESC lines (named VUB-hESC) in our laboratory.