Rapid isolation of human CD34 hematopoietic stem cells--purging of human tumor cells

Transplantation. 1992 May;53(5):1011-9. doi: 10.1097/00007890-199205000-00009.

Abstract

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.

MeSH terms

  • Antibodies, Monoclonal
  • Antigens, CD / analysis*
  • Antigens, CD / genetics
  • Antigens, CD34
  • Bone Marrow Cells
  • Breast Neoplasms / pathology
  • Carcinoma, Small Cell / pathology
  • Cell Adhesion
  • Cell Separation / methods*
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / immunology*
  • Humans
  • Immunoglobulin G / analysis
  • Lectins / metabolism
  • Leukemia, Myeloid, Acute / pathology
  • Neuroblastoma / pathology
  • Phenotype
  • Plant Lectins*
  • Soybean Proteins*
  • Stem Cells / immunology

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, CD34
  • Immunoglobulin G
  • Lectins
  • Plant Lectins
  • Soybean Proteins
  • soybean lectin