Growth of human mast cells from bone marrow and peripheral blood-derived CD34(+) pluripotent hematopoietic cells.
Human mast cells (HuMCs) are derived from CD34(+) pluripotent hematopoietic cells which are KIT (CD117)(+) and FcεRI(-), and lack lineage-specific 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 specific 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 first week). Weekly hemidepletions and removal of adherent cells and/or debris enables the investigator to obtain HuMC cultures, identified by Wright-Giemsa and acidic toluidine blue stains, by 8-10 weeks.
Laboratory of Allergic Diseases, Mast Cell Biology Sections, NIH/National Institute of Allergy and Infectious Diseases, Bethesda, MD, USA.
Geethani
G
Bandara
Dean D
DD
Metcalfe
Arnold S
AS
Kirshenbaum
D018952:Antigens, CD34 / Q000378:metabolism*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D015925:Cryopreservation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D008407:Mast Cells / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D013194:Staining and Labeling
D018952:Antigens, CD34