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- Title
Periferik kandan izole edilen hematopoietik kök hücrelerin megakaryosite farklılaştırılması.
- Authors
Bingöl Özakpınar, Özlem; Maurer, Anne-marie; Adıgüzel, Cafer; Tuğçe Çilingir, Özlem; Demir, Muzaffer; Uras, Fikriye
- Abstract
Objective: Thrombocytopenia remains a serious problem in patients treated with highdose chemotherapy and bone marrow transplantation. In recent years, infusion of ex vivo expanded megakaryocytes (Mk) progenitors into patients has been proposed as a strategy for shortening the time of platelet engraftment. The development of in vitro culture methods to obtain sufficient numbers of Mks from haematopoietic stem cells (HSC) is an important target in basic and clinical research projects. The aim of this study was to develop a two-step ex vivo expansion culture system of Mk progenitors from peripheral blood stem cells (PBSC). Methods: PBSC were harvested from three healthy adult donors. CD34+ cells were isolated and cultured in serum free media supplemented with thrombopoietin (TPO) (50 ng/ml), Interleukin 3 (IL-3) (20 ng/ml) and Interleukin 6 (IL-6) (20 ng/ml) for 12 days followed by an incubation with IL-6 (20 ng/ml) and TPO (50 ng/ml) for another 9 days. The differentiation of Mks was monitored by flow cytometry (% of CD34+/41+ cells). The morphology of the cells was studied by light, electron and fluorescence microscopy. Results: Morphological analysis of cells generated after 7 days of culture showed typical aspects of developing Mks. The percentage of CD41+ cells was higher than 70 on day 21. Conclusion: The results obtained in this study demonstrated that this two-step culture system is an effective method to obtain high rates of megakaryocytes. It is obvious that this promising method needs further development for clinical applications.
- Subjects
THROMBOCYTOPENIA; CANCER chemotherapy; BONE marrow transplantation; MEGAKARYOCYTES; HEMATOPOIETIC stem cells; STEM cell research
- Publication
Turkish Journal of Biochemistry / Turk Biyokimya Dergisi, 2013, Vol 38, Issue 3, p243
- ISSN
0250-4685
- Publication type
Article
- DOI
10.5505/tjb.2013.52714