Irfan Ahmed Siddiqui
Research Scholar, CMJ University, Shillong, Meghalaya
Dr. K. Babu Rao
Principal Donbasco College Pharmacy, Guntur District, AP
Download PDFCD34+ cells, present at a frequency of 0.18 ± 0.052% among leukocytes from peripheral blood (PB), can be rapidly and efficiently en- riched to a frequency of 38.6-87.1% (&4*4 by high-gradient magnetic cell separation (MACS) for Immunopheno- typing, characterization in colony-forming cell assays, and further purification to homogeneity (>98%) by multiparameter fluorescence-activated cell sorting (FACS). Enriching PB-CD34+ cells for immunophenotyping allows the detection of small subpopulations, expressing the B-cell antigens CD10, CD19, and CD20, the T-cell antigens CD45RA and CD7, and a small subpopulation expressing high levels of CD34 which mostly coexpress CD19 CD20 ; and CD38 . All PB-CD34 + cells express elevated levels of CD71 (transfer-rin receptor), with a subpopulation of high expressing cells, and CD38. Some cells express CD33. MACSenriched PB-CD34+ cells show "normal" hematopoie-tic colony formation in vitro. The ease and efficiency of purification of large numbers of CD34+ cells from PB by MACS is not only relevant for the characterization of migrating stem cells but also opens new possibilities for stem cell transplantation & genetic manipulation of the hematopoietic system.
Keywords: Migrating stem cells, flow cytometry, cell sorting, fluorescence-activated cell sorting, immunophenotype, colony-forming assay
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