Flow cytometry-based characterization of label-retaining stem cells following transplacental BrdU labelling

• DOI: 10.1042/cb20100335
• Publication Date: 2011-01-10T21:50:00Z
• Authors: Shiv Poojan; Sushil Kumar
• Keywords: Characterization; Transplacental

Abstract A method to characterize and culture stem cells from neonate mouse epidermis after transplacental BrdU (bromo-deoxyuridine) administration is described. We have characterized by their properties viz. retain label, adhere rapidly onto collagen—fibronectin substratum express a specific biomarker beta-1-integrin. BrdU-labelled (detected using monoclonal antibody) constituted sum of 18% the total number cells. The ability freshly isolated keratinocytes [LRCs (label-retaining cells)] bind primary antibody or pick up PI (propidium iodide) stain was distinguishable. Viable LRCs did not PI. Such cells, termed EpSC (epidermis cell), were negative positive. 6% cell yield. Culture in low Ca2+ medium susceptibility differentiation presence high levels further This protocol useful for studying carcinogenesis.