Multicentre European study comparing selection techniques for the isolation of CD34+ cells.
Primitive haemopoietic cells are required for studies in both the clinical and research fields and a number of systems have been developed to facilitate isolation of these haemopoietic cell populations. We have analysed the results from several European centres using positive selection of CD34+ cells from haemopoietic tissues (n = 110). Four selection techniques including immunoaffinity columns (Ceprate LC), immunomagnetic beads (Dynabeads, Baxter Isolex 50) and submicroscopic magnetic beads (MACS) were used and the selected CD34+ cells were assessed for purity, yield and enrichment of colony-forming cells (CFC). The mean purities for all samples ranged from 68.4-78.4% for MACS, 33.9-69.9% for Dynabeads, 46.9-66.8% for Ceprate LC and 43.2-65% for Baxter Isolex 50. Yields were variable with all techniques. On average CFC enrichment using the immunoaffinity columns was greater than that observed for the other systems. Some techniques appear to be problematic and may require further expertise to improve the results. Nevertheless, the study demonstrates that highly purified CD34+ cells can be isolated from various haemopoietic sources, though yield and CFC enrichment varies significantly depending on the technique selected. This extends our previous report indicating that not all selection methods generate similar results and that there are differences in the purity, number and colony-forming ability of the cells recovered.
de Wynter
E A
EA
Paterson Institute for Cancer Research, Withington, Manchester, UK.
Ryder
D
D
Lanza
F
F
Nadali
G
G
Johnsen
H
H
Denning-Kendall
P
P
Thing-Mortensen
B
B
Silvestri
F
F
Testa
N G
NG
D018952:Antigens, CD34 / Q000032:analysis*; D001781:Blood Component Removal; D002469:Cell Separation / Q000379:methods*; D006412:Hematopoietic Stem Cells; D006801:Humans; D018189:Immunomagnetic Separation
D018952:Antigens, CD34