Growth of human mast cells from bone marrow and peripheral blood-derived CD34+ pluripotent hematopoietic cells

Abstract

Human mast cells (HuMCs) are derived from CD34 + pluripotent hematopoietic cells which are KIT (CD117) + and FcεRI -, and lack lineage-specifi c surface markers. Bone marrow and peripheral blood are the two readily available sources for obtaining CD34 + cells from which HuMCs can be cultured. CD34 + cells are isolated and enriched by magnetic separation columns and stored under specifi c conditions until ready for use. Alternatively, enriched CD34 + cells may be immediately cultured in serum-free culture media containing recombinant human (rh) stem cell factor (SCF), rhIL-6, and rhIL-3 (added only during the fi rst week). Weekly hemidepletions and removal of adherent cells and/or debris enables the investigator to obtain HuMC cultures, identifi ed by Wright-Giemsa and acidic toluidine blue stains, by 8–10 weeks.