In this video we demonstrate how our lab routinely passages HuES human embryonic stem cell lines with trypsin. Brought to you by JoVE.
General
We recommend thawing no more than one sample at a given time to ensure easy handling. The handler should take care not to leave cultures at room temperature and low CO2 for long periods of time. All centrifugation of live cells is done at 500-600 x g for 5 min at room temperature.
MEFs (Mouse Embryonic Feeder Cells)
In this video we demonstrated how we routinely passage HuES human embryonic stem cell lines in our laboratory. Efficient and regular splitting and passaging is essential for maintaining a healthy human embryonic stem cell line. Trypsin mediated passage is a significant advantage of HuES cell lines, however, it is important not to over trypsinize cultures or to have a large proportion of single cells during re-platting.