Separation of Lectin-Binding Cells Using Polystyrene Culture Devices with Covalently Immobilized Soybean Agglutinin

Abstract

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.

In vitro protection of umbilical cord blood–derived primitive hematopoietic stem progenitor cell pool by mannose‐specific lectins via antioxidant mechanisms

“…Lectins have been frequently used as cell separation agents for enrichment or isolation of a particular cell type from a heterogeneous cell population, including hematopoietic cells, due to their selective binding on the carbohydrate residues that are expressed on the cell surface as a function of developmental or differentiation stage. Soybean agglutinin has been used for negative selection of HSPCs from human marrow cells 33 as well as for T‐cell depletion in BM allografts 34,35 . Wheat germ agglutinin lectin has been used to enrich murine HSCs 36‐38 .…”

Selection of hematopoietic stem cells with a combination of galactose‐bound vinyl polymer and soybean agglutinin, a galactose‐specific lectin

“…This approach has facilitated successful marrow transplantation by expanding hematopoietic stem cells and reducing graft‐versus‐host disease (GVHD) 9 . Other trials have utilized a purging method with polystyrene culture devices carrying covalently immobilized SBA 10 …”