Identification, isolation, quantification and systems approach towards CD34, a biomarker present in the progenitor/stem cells from diverse lineages

•We illustrate methodology to Identify, isolate and quantify CD34+ progenitor/stem cells from three diverse lineages.•Understanding the importance of CD34 positive cells in translational medicine is discussed.•Identifications of materials required are discussed.•The difficulties (do and dont’s) involved in the methodology are discussed. Mesenchymal stem cells (MSCs) constitute the diverse progenitor populations in almost every tissue and are of immense importance in the field of regenerative medicine. CD34 is a cell surface glycoprotein identified first as a marker for the MSCs of hematopoietic origin. CD34 is now known to be expressed in cells of diverse lineages (tissues of non-hematopoietic origin) such as ectoderm, mesoderm and endoderm and is considered as a general marker for progenitor cells. Here, we present detailed protocols to obtain pure populations of MSCs from three diverse lineages such as skeletal muscle, skin, and liver from mouse tissues. We also present here the protocol for systems biology approach (proteomic analysis) of these purified cells. This proteomic approach can elucidate key signalling pathways and proteins utilized by these CD34 positive cells in undifferentiated and differentiated conditions. Furthermore in-depth proteomic analysis can also identify the altered proteome which is responsible for their function during non-clinical and clinical conditions.