Purification and partial characterization of a human hematopoietic precursor population.
This study reports the development of an assay, the Pre-colony-forming unit (CFU) assay, which detects human hematopoietic precursors. The Pre-CFU assay is based on the observation that precursors to CFU-granulocyte-macrophage (CFU-GM) that are undetectable in clonogenic assays differentiate into CFU-GM preferentially following treatment in suspension culture with recombinant human interleukin-1 alpha (rhIL-1 alpha) combined with rhIL-3. Using the Pre-CFU assay, hematopoietic precursors were detected in human bone marrow depleted of CFU-GM progenitors and differentiated hematopoietic elements via 4-hydroperoxycyclophosphamide treatment coupled with selection for CD34+ cells (4-HCresistant/CD34+ marrow). Additionally, the Pre-CFU assay detected recovery of hematopoiesis substantially earlier than the CFU-GM assay in primates following myeloablation with 5-fluorouracil. The Pre-CFU assay was used to asses purification of a phenotypically defined hematopoietic precursor population, the lin-CD34+ population. The lin-CD34+ population lacks detectable surface markers for T-cell, B-cell, natural killer cell, and myeloid lineage, possesses the CD34 antigen, is devoid of CFU-GM progenitors, and yields Pre-CFU assay values comparable with 4-HCresistant/CD34+ marrow. Using a combination of phenotypic analysis and Pre-CFU assay analysis, the action of rhIL-1 alpha plus rhIL-3 treatment on lin-CD34+ cells was further characterized. The data indicate that rhIL-1 alpha plus rhIL-3 treatment induces proliferation and differentiation of early hematopoietic precursors into progenitors and terminally differentiated cells, without inducing a significant expansion of the precursor population itself.
Smith
C
C
Bone Marrow Transplant Service, Memorial Sloan Kettering Cancer Center, New York, NY 10021.
Gasparetto
C
C
Collins
N
N
Gillio
A
A
Muench
M O
MO
O'Reilly
R J
RJ
Moore
M A
MA
D000818:Animals; D018952:Antigens, CD34; D000943:Antigens, Differentiation / Q000276:immunology; D001853:Bone Marrow / Q000276:immunology / Q000502:physiology; D001854:Bone Marrow Cells; D002455:Cell Division; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay / Q000379:methods*; D003520:Cyclophosphamide / Q000031:analogs & derivatives / Q000494:pharmacology; D005472:Fluorouracil / Q000494:pharmacology; D006098:Granulocytes / Q000166:cytology / Q000502:physiology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D008251:Macaca; D008264:Macrophages / Q000166:cytology / Q000502:physiology; D008297:Male; D010641:Phenotype; D011994:Recombinant Proteins / Q000494:pharmacology
D018952:Antigens, CD34; D000943:Antigens, Differentiation; D007377:Interleukin-3; D011994:Recombinant Proteins; C012358:4-hydroxycyclophosphamide; D003520:Cyclophosphamide; D005472:Fluorouracil