Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Schwinger, W; Benesch, M; Lackner, H; Kerbl, R; Walcher, M; Urban, C.
Comparison of different methods for separation and ex vivo expansion of cord blood progenitor cells.
Ann Hematol. 1999; 78(8):364-370
Doi: 10.1007%2Fs002770050530
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- Führende Autor*innen der Med Uni Graz
- Schwinger Wolfgang
- Co-Autor*innen der Med Uni Graz
- Benesch Martin
- Kerbl Reinhold
- Lackner Herwig
- Urban Ernst-Christian
- Walcher Wolfgang
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- Abstract:
- Umbilical cord blood is capable of hematopoietic stem cell reconstitution in children. However, the major limitation of cord blood is a relatively low content of pluripotent progenitor cells. Thus, safe engraftment for adolescents and for adults is still not predictable and a technology for ex vivo expansion of umbilical cord blood cells is desirable. In a first step, four different methods of red cell depletion followed by magnetic cell sorting of CD34+ cells were evaluated in this study in order to assess the efficacy and safety of optimal stem cell recovery. A modified two-step Ficoll gradient separation and a hydroxyethyl starch separation tended to produce a better WBC/MNC recovery (median 94.2+/-2.44% vs. 90.2+/-5. 8%) as compared with standard Ficoll gradient separation and a gelatin-based procedure (median 78.35+/-7.1% vs. 67.2+/-5.5%). However, the recovery of CD34+ cells after magnetic cell sorting did not reach a statistically significant difference after the four different methods of red cell depletion, indicating that the recovery of WBC/MNC is not predictably correlated with the recovery of stem cells within these fractions. In a second step, we established three different cytokine combinations by adding the megakaryocyte growth and development factor +/- erythropoietin and granulocyte colony-stimulating factor to a fetal calf serum containing medium with Flt 3, stem cell factor, and interleukin-3. Net expansion of total colony-forming cells 20- to 50-fold and expansion of colony-forming cells after 5 weeks of culture 1.5- to 3-fold were obtained over a period of 7-14 days. These results demonstrate that cord blood stem cells can be expanded substantially in this short-term culture system.
- Find related publications in this database (using NLM MeSH Indexing)
- Antigens, CD34 - blood
- Blood Component Removal - methods
- Cell Separation - methods
- Colony-Forming Units Assay - methods
- Cytapheresis - methods
- Erythrocyte Transfusion - methods
- Erythroid Progenitor Cells - cytology
- Female - cytology
- Fetal Blood - cytology
- Granulocytes - cytology
- Hematopoietic Stem Cells - cytology
- Humans - cytology
- Macrophages - cytology
- Megakaryocytes - cytology
- Monocytes - cytology
- Pregnancy - cytology
- Find related publications in this database (Keywords)
- Selection
- Purification
- Expansion
- Cord Blood Stem Cells