Separation of lectin-binding cells using polystyrene culture devices with covalently immobilized soybean agglutinin.
The plant lectin, soybean agglutinin (SBA), has been widely used to separate heterogeneous populations of cells. In the field of bone marrow transplantation, SBA has been used for partial depletion of T cells from bone marrow allografts to reduce graft-vs.-host disease. SBA's high affinity for many different tumor cells has also indicated its use as a tumor purging agent for autologous bone marrow transplants. We have compared two methods of cell separation using either soluble SBA agglutination, or SBA covalently attached to an activated polystyrene surface. The nonbinding SBA-cell populations generated by these two procedures were very similar in terms of cell recovery, light scatter properties, and phenotypic profile. Notably, both SBA- fractions were enriched in cells with the known progenitor markers, CD34, CD33, and HLA-DR, and were relatively depleted of SBA binding cells. In addition, the activity of each SBA- cell population was measured in vitro in short-term progenitor assays. Here, both SBA- populations were significantly enriched for CFU-GM. When device-separated SBA- cell populations were seeded into long-term bone marrow culture, they produced both increased progenitor activity and cell proliferation compared to unseparated BMMCs. The polystyrene technology described here could reduce or eliminate many of the drawbacks of soluble SBA agglutination, making SBA cell separation a viable and convenient technique for clinical application.
Schain
L R
LR
Applied Immune Sciences, Inc., Santa Clara, CA 95054.
Okrongly
D
D
Okarma
T B
TB
Lebkowski
J S
JS
D015703:Antigens, CD / Q000032:analysis; D001853:Bone Marrow / Q000276:immunology; D001854:Bone Marrow Cells; D002469:Cell Separation / Q000295:instrumentation*; D048429:Cell Size; D002478:Cells, Cultured; D006410:Hematopoiesis / Q000502:physiology; D006801:Humans; D037102:Lectins / Q000378:metabolism; D010641:Phenotype; D037121:Plant Lectins; D011137:Polystyrenes; D011485:Protein Binding; D012016:Reference Values; D030262:Soybean Proteins; D013025:Glycine max; D010835:Phytohemagglutinins
D015703:Antigens, CD; D037102:Lectins; D037121:Plant Lectins; D011137:Polystyrenes; D030262:Soybean Proteins; C028824:soybean lectin; D010835:Phytohemagglutinins