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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۳۶-۳۶
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عنوان انگلیسی Ps-38: Effective Megakaryocytic Differentiation of Umbilical Cord Blood Hematopoietic Stem/Progenitor Cells Using A Two-Phase Protocol
چکیده انگلیسی مقاله Objective: There is a high need for the ex-vivo expansion of Hematopoietic stem and progenitor cells (HSC/HPC), especially those from the umbilical cord blood (UCB) which are limited in number, for their extensive use in hematopoietic transplantation. Likewise, considering that platelets do not withstand refrigeration and having short lifetime of 5 days, a high demand for cells with megakaryocytic potential exists aiming at the clinical scale production of platelets for transfusion. However the complexity of hematopoiesis, the heterogeneity of culture population and the different combination of culture parameters has impaired to establish an efficient protocol to be used for platelet production from UCB. Thus, we aimed to develop a two-phase ex-vivo protocol for expansion and differentiation of HSC/HPC from UCB towards megakaryocyte lineage using a static co-culture system. Materials and Methods: HSC/HPC from UCB were cocultured with mesenchymal stem cells from bone marrow (BM-MSC) in a serum free medium with a cytokine cocktail including TPO, SCF, FLT3, b-FGF (50, 60, 55 and 5 ng/mL, respectively). After 7 days of expansion, the hematopoietic cells were retrieved and submitted to a differentiation protocol using IMDM medium supplemented with 10% fetal bovine serum and a cocktail of TPO and IL3 (100 and 10 ng/mL, respectively). Results: HSC/HPC were co-cultured with BM-MSC for 7 days, reaching a 20 ± 5 fold increase in CD34+ cell number. By the end of differentiation phase, more than 35 % of cells population expressed megakaryocyte lineage surface maker (CD41+) and we were able to produce 50 Megakaryocytes per input CD34+ cells. Inverted and electron microscopy images confirmed megakaryocyte and platelet morphology. Conclusion: HSC/HPC from UCB were successfully expanded in a static co-culture system, phenotype and morphological analysis suggested effective differentiation of HSC/HPC towards the megakaryocyte lineage.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/843
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