Chen Te-Wei, Yao Chao-Ling, Chu I-Ming, Chuang Tzu-Lin, Hsieh Tzu-Bou, Hwang Shiaw-Min
Bioresource Collection and Research Center, Food Industry Research and Development Institute, 331 Shih-Pin Road, Hsinchu 30062, Taiwan.
Biochem Biophys Res Commun. 2009 Jan 2;378(1):112-7. doi: 10.1016/j.bbrc.2008.11.019. Epub 2008 Nov 21.
Ex vivo generation of megakaryocytes from hematopoietic stem cells (HSCs) is crucial to HSC research and has important clinical potential for thrombocytopenia patients to rapid platelet reconstruction. In this study, factorial design and steepest ascent method were used to screen and optimize the effective cytokines (10.2 ng/ml TPO, 4.3 ng/ml IL-3, 15.0 ng/ml SCF, 5.6 ng/ml IL-6, 2.8 ng/ml FL, 2.8 ng/ml IL-9, and 2.8 ng/ml GM-CSF) in megakaryocyte induction medium that facilitate ex vivo megakaryopoiesis from CD34(+) cells. After induction, the maximum fold expansion for accumulated megakaryocytes was almost 5000-fold, and the induced megakaryocytes were characterized by analysis of gene expression, polyploidy and platelet activation ability. Furthermore, the combination of megakaryocyte induction medium and HSC expansion medium can induce and expand a large amount of functional megakaryocytes efficiently, and might be a promising source of megakaryocytes and platelets for cell therapy in the future.
从造血干细胞（HSCs）体外生成巨核细胞对于HSC研究至关重要，并且对于血小板减少症患者快速重建血小板具有重要的临床潜力。在本研究中，采用析因设计和最速上升法筛选并优化了巨核细胞诱导培养基中的有效细胞因子（10.2 ng/ml血小板生成素、4.3 ng/ml白细胞介素-3、15.0 ng/ml干细胞因子、5.6 ng/ml白细胞介素-6、2.8 ng/ml促血小板生成素、2.8 ng/ml白细胞介素-9和2.8 ng/ml粒细胞-巨噬细胞集落刺激因子），这些细胞因子有助于从CD34(+)细胞体外生成巨核细胞。诱导后，累积巨核细胞的最大扩增倍数近5000倍，并通过基因表达分析、多倍体分析和血小板激活能力分析对诱导生成的巨核细胞进行了表征。此外，巨核细胞诱导培养基和HSC扩增培养基的组合能够高效诱导并扩增大量功能性巨核细胞，可能成为未来细胞治疗中巨核细胞和血小板的一个有前景的来源。
