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Updated: Jun 20, 2026
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Isolation Method for Long-Term and Short-Term Hematopoietic Stem Cells
Published on: May 19, 2023
Hematopoietic stem cell separation for experimental purposes--methodic limitations.
Pavla Luzna1, Dana Kylarova, Martin Novak
1Department of Histology and Embryology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, 775 15, Czech Republic. pavla83@hotmail.com
Summary
Immunomagnetic separation of CD117+ cells from bone marrow is not effective for research. Cell suspension staining is a viable method for examining immature blood cell morphology and immunophenotype.
Area of Science:
- Hematology
- Immunology
- Cell Biology
Background:
- Protooncogene CD117 is a crucial cytokine receptor for hematopoietic development.
- Current methods for isolating sufficient quantities of CD117+ bone marrow cells for research are inadequate.
Purpose of the Study:
- To develop an immunomagnetic separation technique for isolating CD117+ hematopoietic stem cells.
- To assess the expression of BCL-2 family proteins in these isolated cells.
Main Methods:
- Analyzed 120 human and murine bone marrow samples using magnetic separation.
- Employed indirect fluorescent immunocytochemistry to stain cells for CD117, BCL-2, BAX, and CD33.
Main Results:
- Flow cytometry revealed low CD117+ cell yields (2.6%), insufficient for experiments.
- Cell suspension staining identified CD117 in 70% of CD33+ cells in chronic myeloid leukemia.
- BCL-2 and BAX expression was found in 80% and 5% of cells, respectively.
Conclusions:
- CD117 immunomagnetic separation is unsuitable for experimental bone marrow cell isolation.
- Cell suspension staining is the preferred method for bone marrow cell examination, revealing surface antigens and cytoplasmic proteins.
