Comparison of Enzymatic and Non-Enzymatic Means of Dissociating Adherent Monolayers of Mesenchymal Stem Cells

• Main Authors: Heng Boon, Cowan Catherine, Basu Shubhayu
• Format: Article
• Language: English
• Published: BMC 2009-01-01
• Series: Biological Procedures Online
• Subjects: Dissociation; Enzyme; Mesenchymal; Stem cells; Trypsin
• Online Access: http://www.biologicalproceduresonline.com/content/11/1/161

<p>Abstract</p> <p>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&#8211;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.</p>