An eight-fold ex vivo expansion of long-term culture-initiating cells from umbilical cord blood in stirred suspension cultures.
Simultaneous ex vivo expansion of different progenitor cell types may be beneficial for cord blood (CB) transplantation, to overcome a potential limitation due to restricted cell numbers. Therefore, 1.5 x 10(6) CD34+ cells isolated from fresh or thawed CB samples were inoculated in a large-scale stirred suspension bioreactor and cultured in the presence of Flt3-L, SCF and IL-3. At days 0, 7, 10, 14, 21 and 28, the spinner cultures were analyzed for viable cells, colony-forming cells (CFC), including erythroid burst-forming unit (BFU-E), granulocyte-macrophage colony-forming unit (CFU-GM) and granulocyte-erythrocyte-megakaryocyte-monocyte colony-forming unit (CFU-GEMM) as well as long-term culture-initiating cells (LTC-IC). Expansion of thawed CD34+ cells resulted in a substantial amplification of total cells (maximal at day 28: 154 +/- 132-fold), CFC (maximal at day 14: 45 +/- 36-fold), CFU-GM (maximal at day 14: 88 +/- 85-fold), CFU-GEMM (maximal at day 7: 4 +/- 2-fold) and of LTC-IC (maximal at day 10: 8 +/- 3-fold). There was no significant difference between fresh and thawed CD34+ cells. These results demonstrate that simultaneously committed progenitors as well as the more immature CFU-GEMM and LTC-IC can be substantially amplified from CD34+-enriched CB samples in large-scale stirred suspension cultures within 7-14 days without exhausting the proliferative potential and, thus, it may be possible to improve CB transplantation by ex vivo generated cells.
Bone Marrow Donor Center with Eurocord Bank Germany and Transplantation Immunology, Heinrich Heine University, Düsseldorf.
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G
Kögler
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J
Callejas
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RV
Sorg
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P
Wernet
D018952:Antigens, CD34; D001772:Blood Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D019650:Hematopoietic Stem Cell Mobilization; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans
D018952:Antigens, CD34