I want to transition my iPSCs from MEFs to Matrigel. What is the best method for transition for the cells and is there anything I should add to the media?
This question is part of the following Ask The Expert session:
Answered by:
Answer
Instructions for transitioning to Essential 8 medium on Geltrex, which is comparable to Matrigel, can be found here: http://www.lifetechnologies.com/us/en/home/references/protocols/cell-culture/stem-cell-protocols/ipsc-protocols/culturing-puripotent-stem-cells-essential-8-medium.html under the “Adaptation using Geltrex® LDEV-Free, hESC-Qualified Basement Membrane Matrix as an Intermediary” section. In your cause you would be staying on the richer media substrate rather than moving to a leaner matrix such as rhVTN-N. Preliminary experiments suggest that addition of RevitaCell™ Supplement (Cat #A26445-01) to the culture medium during the first 24 hours post-transition can also aid in maintenance of cell health during the transition period.
This post is sponsored by Thermo Fisher Scientific, Inc..