Which passaging method do you recommend for pluripotent stem cells?

All common passaging techniques will work. Our customers use StemMACS iPS-Brew XF, human in combination with traditional colony cutting, clump-splitting, as well as enzymatic single-cell passaging techniques.

At Miltenyi Biotec, we prefer to use StemMACS™ Passaging Solution XF, because it is a very quick and easy method: Remove the old culture media and wash once with PBS. Then, simply add StemMACS Passaging Solution XF for 4 minutes and the colony edges will start to lift off. After removing the solution, use fresh media to dissociate your colonies into clumps and transfer to a new dish.

To enhance the attachment of new PSC lines, we recommend including a Rho-associated protein kinase (ROCK) inhibitor such as StemMACS Y27632 (10 µM final concentration) or StemMACS Thiazovivin (2 µM final concentration) for the plating. The ROCK inhibitor can be removed with the next media change.