Authors: B. Roda, G. Lanzoni, F. Alviano, A. Zattoni, R. Costa, A. Di Carlo, C. Marchionni, M. Franchina, F. Ricci, P. Tazzari, P. Pagliaro, S.Z. Scalinci, L. Bonsi, P. Reschiglian, G.P. Bagnara
Affilation: University of Bologna, Italy
Pages: 195 - 198
Keywords: tag-less cell sorting, stem cell, flow separation
New methods for stem cells isolation are being developed. Common procedures for stem cell sorting involve flow cytometry which makes use of fluorescent or magnetic tags to isolate and separate cells which are phenotypically different. However, immuno-selection is less precise and easy because of the lack of specific stem cell marker for several stem cell populations; cell-type-specific markers are limited and often recognize multiple members of a lineage and immuno-tags for cell identification are known to affect cell functionality and recovery. Among stem cells, mesenchymal stem cells (MSCs) are promising candidates for cell therapy applications. This work presents a fractionation protocol based on a proprietary technology for the tag-less, flow-assisted purification, characterization and sorting of MSCs. The fractionation process is based on differences in cell biophysical features and it allows to preserve cell viability and features. The protocol can be applied to isolate and separate MSCs from potential phenotypically different contaminants when cells are isolated from clinical specimens in order to reduce the number of cell culture passages for MSC selection; to distinguish MSCs derived from different sources, and finally to sort stem cells from an MSC population isolated from a single source, obtaining the highest differentiation yield.