Rapid isolation of human CD34 hematopoietic stem cells--purging of human tumor cells.
Human CD34+ hematopoietic stem cells were purified using a new technology in which monoclonal antibodies are covalently immobilized on polystyrene surfaces. The CD34+ cell isolation scheme involved three sequential processes: (1) purification of bone marrow mononuclear cells; (2) enrichment of CD34+ cells using covalently immobilized soybean agglutinin; and (3) positive selection of CD34+ cells using polystyrene surfaces coated with the anti-CD34 monoclonal antibody ICH3. CD34+ cells purified by this process have both low-to-medium forward light scatter and low 90 degrees light-scatter properties. Moreover, the purified CD34+ cells are greater than 85% viable, express appropriate characteristic surface antigens, and are 10-50-fold enriched in short- and long-term hematopoietic activity. CD34+ cells collected in this manner from bone marrow samples contaminated with radiolabeled breast carcinoma, neuroblastoma, acute myelogenous leukemia, or small cell lung carcinoma cells were 99.9% depleted of the tumor cells. The CD34+ cell selection devices are sterile and are easily scaled-up to process clinical scale bone marrow samples.
Lebkowski
J S
JS
Applied Immune Sciences, Inc., Menlo Park, California 94025.
Schain
L R
LR
Okrongly
D
D
Levinsky
R
R
Harvey
M J
MJ
Okarma
T B
TB
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis* / Q000235:genetics; D018952:Antigens, CD34; D001854:Bone Marrow Cells; D001943:Breast Neoplasms / Q000473:pathology; D018288:Carcinoma, Small Cell / Q000473:pathology; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D037102:Lectins / Q000378:metabolism; D015470:Leukemia, Myeloid, Acute / Q000473:pathology; D009447:Neuroblastoma / Q000473:pathology; D010641:Phenotype; D037121:Plant Lectins; D030262:Soybean Proteins; D013234:Stem Cells / Q000276:immunology; D010835:Phytohemagglutinins
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018952:Antigens, CD34; D007074:Immunoglobulin G; D037102:Lectins; D037121:Plant Lectins; D030262:Soybean Proteins; C028824:soybean lectin; D010835:Phytohemagglutinins