Large generation of megakaryocytes from serum-free expanded human CD34+ cells.
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.
Chen
Te-Wei
TW
Bioresource Collection and Research Center, Food Industry Research and Development Institute, 331 Shih-Pin Road, Hsinchu 30062, Taiwan.
Yao
Chao-Ling
CL
Chu
I-Ming
IM
Chuang
Tzu-Lin
TL
Hsieh
Tzu-Bou
TB
Hwang
Shiaw-Min
SM
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
D018952:Antigens, CD34; D016895:Culture Media, Serum-Free; D016207:Cytokines; D011994:Recombinant Proteins