title: Large generation of megakaryocytes from serum-free expanded human CD34+ cells.
abstract: 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.
authors.affiliation: Bioresource Collection and Research Center, Food Industry Research and Development Institute, 331 Shih-Pin Road, Hsinchu 30062, Taiwan.
authors.forename: Te-Wei
authors.initials: TW
authors.lastname: Chen
authors.forename: Chao-Ling
authors.initials: CL
authors.lastname: Yao
authors.forename: I-Ming
authors.initials: IM
authors.lastname: Chu
authors.forename: Tzu-Lin
authors.initials: TL
authors.lastname: Chuang
authors.forename: Tzu-Bou
authors.initials: TB
authors.lastname: Hsieh
authors.forename: Shiaw-Min
authors.initials: SM
authors.lastname: Hwang
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D016895:Culture Media, Serum-Free / Q000737:chemistry / Q000494:pharmacology; D016207:Cytokines / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology* / Q000378:metabolism; D015539:Platelet Activation; D011003:Ploidies; D011994:Recombinant Proteins / Q000494:pharmacology
chemical_list: D018952:Antigens, CD34; D016895:Culture Media, Serum-Free; D016207:Cytokines; D011994:Recombinant Proteins