Comparison of two immunomagnetic separation technologies to deplete T cells from human blood samples.
The objective of this study was to compare the performance of two immunomagnetic separation technologies to deplete T cells from buffy coats of human blood. Specifically, two versions of the commercial MACS(R) Technology: MiniMACS and SuperMACS, and a prototype, flow-through system, the QMS, were evaluated. Peripheral blood mononuclear leukocytes (PBL) were isolated from buffy coats and an immunomagnetic separation of CD3(+) cells was conducted using company and optimized labeling protocols. To mimic peripheral blood containing bone marrow purged hematopoietic stem cells, HSC, CD34 expressing-cells (KG1a) were spiked into PBL prior to T-cell depletion once optimized depletion conditions were determined. Once the labeling protocol was optimized, the MiniMACS system performed well by producing a highly enriched CD3(+) fraction, and a respectable level of depletion of T cells and recovery of KG1a cells in the depleted fraction; an average log(10) depletion of T cells of 2.88 +/- 0.17 and an average recovery of the KG1a cells of 60.8 +/- 5.94% (n = 14). The performance of the SuperMACS system was very similar with an average log(10) depletion of T cells of 2.89 +/- 0.22 and an average recovery of KG1a of 63.1 +/- 8.55% (n = 10). In contrast, the QMS system produced an average log(10) depletion of T cells of 3.98 +/- 0.33 (n = 16) with a corresponding average recovery of 57.9 +/- 16.6% of the spiked CD34+ cells. The aforementioned QMS performance values were obtained using sorting speeds ranging from 2.5 x 10(4) to 1.7 x 10(5) cells per second. It is suggested that the lack of a 100% recovery of the unlabeled KG1a cells is the result of a previously reported "drafting" phenomena which pulls unlabeled cells in the direction of the magnetically labeled cells thereby resulting in loss of the unlabeled cells.
Lara
Oscar
O
Department of Chemical and Biomolecular Engineering, The Ohio State University, 120 Koffolt Laboratories, 140 W. 19th Avenue, Columbus, 43210, USA.
Tong
Xiaodong
X
Zborowski
Maciej
M
Farag
Sherif S
SS
Chalmers
Jeffrey J
JJ
D018952:Antigens, CD34 / Q000032:analysis; D001773:Blood Cells / Q000166:cytology*; D017252:CD3 Complex / Q000032:analysis; D002478:Cells, Cultured; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D018189:Immunomagnetic Separation / Q000295:instrumentation / Q000379:methods*; D007519:Isoantigens / Q000032:analysis; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D008212:Lymphocyte Depletion; D013601:T-Lymphocytes / Q000166:cytology*
D018952:Antigens, CD34; D017252:CD3 Complex; D007519:Isoantigens; C060224:Kg, red cell antigen