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C-kit positive porcine bone marrow progenitor cells identified and enriched using recombinant stem cell factor
Artur Summerfield1, Michael P Horn, Gabriela Lozano
1Institute of Virology and Immunoprophylaxis, Sensemattstrasse 293, CH-3147 Mittelhäusern, Switzerland. Artur.Summerfield@ivi.admin.ch
Journal of Immunological Methods
|September 16, 2003
Summary
Researchers developed recombinant porcine stem cell factor (SCF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) to identify and isolate porcine hematopoietic progenitor cells. This breakthrough enables detailed porcine hematological studies previously hindered by a lack of specific antibodies.
Area of Science:
- Immunology
- Hematology
- Cell Biology
Background:
- Porcine hematological research is limited by the absence of monoclonal antibodies for CD34 and CD117 on hematopoietic progenitors.
- Accurate identification and isolation of porcine hematopoietic stem and progenitor cells are crucial for advancing research in this field.
Purpose of the Study:
- To describe the enumeration, phenotyping, and isolation of porcine hematopoietic progenitor cells.
- To characterize porcine hematopoietic progenitor cells expressing the stem cell factor receptor (c-kit, CD117).
Main Methods:
- Expression and biological activity assessment of recombinant porcine stem cell factor (rpSCF) and granulocyte-macrophage colony-stimulating factor (rpGM-CSF) using the HEK293 cell system.
- Functional assays including TF-1 cell line proliferation, porcine bone marrow hematopoietic cell (BMHC) proliferation, and dendritic cell generation.
- Immunological detection and binding studies of rpSCF on BMHC using a 6-histidine epitope tag.
- Cell sorting techniques (magnetic and fluorescence-activated) for enrichment of SCF-ligating BMHC.
Main Results:
- Recombinant porcine SCF and GM-CSF were biologically active, inducing proliferation in human TF-1 cells and porcine BMHC.
- rpSCF demonstrated synergistic effects with rpGM-CSF on BMHC proliferation and with GM-CSF/TNF-alpha on dendritic cell generation.
- SCF binding was observed on 4-11% of BMHC, specifically on the SWC3(low/-)SWC8- subset, characteristic of immature progenitors.
- SCF expression was absent on monocytic, granulocytic, and B cell lineages.
- Enrichment of pluripotent progenitor cells was achieved by isolating SCF-ligating BMHC.
Conclusions:
- The development of biologically active recombinant porcine SCF and GM-CSF provides essential tools for porcine hematological research.
- Identification and isolation of porcine hematopoietic progenitor cells expressing c-kit (CD117) are now feasible.
- These advancements will facilitate detailed studies of porcine hematopoiesis, previously limited by antibody availability.