Replating protocol for human induced pluripotent stem cell–derived cardiomyocytes
Künye
Koç, A. ve Çağavi, E. (2022). Replating protocol for human induced pluripotent stem cell–derived cardiomyocytes. Methods in Molecular Biology içinde (161-170. ss.). Humana Press Inc. http://doi.org/10.1007/7651_2021_450Özet
Human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CM) create an unlimited cell source for basic and translational cardiac research. Obtaining hiPSC-CM culture as a single-cell, monolayer or three-dimensional clusters for downstream applications can be challenging. Thus, it is critical to develop replating strategies for hiPSC-CMs by evaluating different enzymatic or nonenzymatic reagents for dissociation and seeding on different coating materials. To reseed hiPSC-CMs with high viability and at structures desirable for the downstream applications, here we defined optimized protocols to dissociate hiPSC-CMs by using collagenase A&B, Collagenase II, TrypLE, and EDTA and reseeding on various matrix materials including fibronectin, laminin, imatrix, Matrigel, and Geltrex. By the replating methods described here, a single cell or cluster-containing hiPSC-CM cultures can be generated effectively.