title: Comparison of enzymatic and non-enzymatic means of dissociating adherent monolayers of mesenchymal stem cells.
abstract: The dissociation of adherent mesenchymal stem cell (MSC) monolayers with trypsin and enzyme-free dissociation buffer was compared. A significantly lower proportion of viable cells were obtained with enzyme-free dissociation buffers compared to trypsin. Subsequently, the dissociated cells were re-seeded on new cell culture dishes and were subjected to the MTT assay 24 h later. The proportion of viable cells that reattached was significantly lower for cells obtained by dissociation with enzyme-free dissociation buffer compared to trypsin. Frozen-thawed MSC displayed a similar trend, yielding consistently higher cell viability and reattachment rates when dissociated with trypsin compared to enzyme-free dissociation buffer. It was also demonstrated that exposure of trypsin-dissociated MSC to enzyme-free dissociation buffer for 1 h had no significant detrimental effect on cell viability.
authors.forename: Boon C
authors.initials: BC
authors.lastname: Heng
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Cowan
authors.forename: Shubhayu
authors.initials: S
authors.lastname: Basu