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In vitro expansion of hematopoietic progenitor cells induces functional expression of Fas antigen (CD95)
- Source :
- Blood. 88:2871-2877
- Publication Year :
- 1996
- Publisher :
- American Society of Hematology, 1996.
-
Abstract
- Fas antigen (Fas Ag; CD95) is a cell surface molecule that can mediate apoptosis. Bcl-2 is a cytoplasmic molecule that prolongs cellular survival by inhibiting apoptosis. To investigate the role of both molecules in hematopoiesis, we evaluated the expression of Fas Ag and Bcl-2 on CD34+ hematopoietic progenitor cells expanded in vitro. CD34+ cells isolated from bone marrow were cultured in iscove's modified Dulbecco's medium supplemented with 10% fetal calf serum, 1% bovine serum albumin, 50 ng/mL stem cell factor, 50 ng/mL interleukin-3 (IL-3), 50 ng/mL IL-6, 100 ng/mL granulocyte colony-stimulating factor, and 3 U/mL erythropoietin for 7 days. Colony-forming unit of granulocytes/macrophages (CFU-GM) and burst-forming unit of erythroids (BFU-E) were expanded 6.9-fold and 8.8-fold in number at day 5 of culture, respectively. Freshly isolated CD34+ cells did not express Fas Ag, whereas approximately half of them expressed Bcl-2. CD34+ cells cultured with hematopoietic growth factors gradually became positive for Fas Ag and rapidly lost Bcl-2 expression. Furthermore, apoptosis was induced in the cultured CD34+ population when anti-Fan antibody (IgM; 1 microgram/mL) was added, as shown by significant decrease in the number of viable cells, morphologic changes, induction of DNA fragmentation, and significant decrease in the number of clonogenic progenitor cells including CFU. GM and BFU-E. These results indicate that functional expression of Fas Ag is induced on CD34+ cells expanded in vitro in the presence of hematopoietic growth factors. Induction of Fas Ag and downregulation of Bcl-2 may be expressed as part of the differentiation program of hematopoietic cells and may be involved in the regulation of hematopoiesis.
- Subjects :
- Adult
medicine.medical_treatment
Immunology
CD34
Apoptosis
Stem cell factor
DNA Fragmentation
Biology
Biochemistry
Colony-Forming Units Assay
medicine
Animals
Humans
fas Receptor
Progenitor cell
Erythropoietin
Cells, Cultured
Interleukin 3
Stem Cell Factor
Growth factor
Antibodies, Monoclonal
Cell Biology
Hematology
Hematopoietic Stem Cells
Fas receptor
Molecular biology
Recombinant Proteins
Hematopoiesis
medicine.anatomical_structure
Gene Expression Regulation
Proto-Oncogene Proteins c-bcl-2
Cattle
Interleukin-3
Bone marrow
Subjects
Details
- ISSN :
- 15280020 and 00064971
- Volume :
- 88
- Database :
- OpenAIRE
- Journal :
- Blood
- Accession number :
- edsair.doi.dedup.....6320f5b751e626a92580242a48ab197b