Isolation of CD34+ hematopoietic progenitor cells in chronic myeloid leukemia by magnetic activated cell sorting (MACS)
Martín-Henao, G.A.; Inglés-Esteve, J.; Cancelas, J.A.; García, J.
Bone Marrow Transplantation 18(3): 603-609
1996
ISSN/ISBN: 0268-3369 PMID: 8879625 Document Number: 457431
We have evaluated an easy and fast immunomagnetic method for positive selection of cells expressing the CD34 antigen from BM, peripheral blood (PB) and apheresis products (AP) of CML patients and healthy adults (HA) in order to further characterize them by immunophenotypic analysis. From an initial frequency of CD34+ cells in the original sample of 1.8 +- 1.7%, CD34+ cells were rapidly and efficiently enriched up to 91.5 +- 6.4% by high-gradient magnetic cell sorting (MACS) (yield 53 +- 21%). A five-dimensional flow cytometric analysis of the immunomagnetic isolated CD34+ cells demonstrated little overlap between CD34+ HLA-DR-lo and CD34+CD38-lo subpopulations in both BM-HA and in BM-CML. Only 16 and 6% of the CD34+HLA-DR-lo and CD34+CD38-lo cells respectively, showed lack of expression of both Ag (CD34+HLA-DR-loCD38-lo) in BMCML samples. Between 60 and 70% of the CD34+ cells expressed the stem cell factor (SCF) receptor (c-KIT, CD117) and there were no differences between BM-HA and BM-CML patients. Moreover, more than 60% of the CD34+HLA-DR-lo cells, co-expressed c-KIT. MACS-enriched BM-CD34+ cells showed normal hematopoietic colony formation in vitro in all the sources analyzed with a higher colony-forming efficiency than the unfractionated sample (MNC).