title: Human adipose tissue-derived stromal cells can differentiate into megakaryocytes and platelets by secreting endogenous thrombopoietin.
abstract: Essentials Manufacturing platelets from a donor-independent source is highlighted in transfusion medicine. We examined the differentiation of adipose tissue-derived stromal cells (ASCs) into platelets. Endogenous thrombopoietin (TPO) induced ASCs differentiation into megakaryocytes and platelets. TPO secretion from ASCs was due to an interaction of transferrin with its receptor CD71.
  
  
  
  Background Ex vivo production of megakaryocytes (MKs) and platelets from a donor-independent source is currently of intense interest in transfusion medicine. Adipose tissue-derived stromal cells (ASCs) constitute an attractive candidate cell source, because inducing these cells into MK lineages requires no gene transfer and only endogenous transcription factors containing p45NF-E2/Maf, an MK-inducing factor. Objectives To examine whether ASCs differentiate into MK lineages by using endogenous thrombopoietin (TPO), a primary cytokine that drives MK lineages. Methods TPO levels were measured by quantitative real-time PCR and ELISA. To investigate the effects of endogenous TPO on MK and platelet production, surface marker expression and functions for platelets were analyzed in ASC-derived cells cultured in the presence or absence of recombinant TPO. Based on a screening test, the role of transferrin receptor CD71 in TPO production and MK differentiation was examined with anti-CD71 antibody, small interfering RNA (siRNA) against CD71 (siRNA-CD71), and CD71-positive/negative cells. Results ASCs secreted TPO during MK differentiation, and the endogenous TPO facilitated MK and platelet production from ASCs. TPO secretion from ASCs occurred in a transferrin-dependent manner. ASCs treated with anti-CD71 antibody or transfected with siRNA-CD71 produced markedly less TPO. The TPO levels and MK yield were significantly higher when CD71-positive ASCs were used than when CD71-negative ASCs were used. Conclusions CD71 might be an appropriate marker for MK progenitor cells among human ASCs, because of the higher capacity of CD71-positive cells to produce TPO and their ability to differentiate into MKs. These findings could help to establish an efficient method for platelet production.
authors.affiliation: Clinical and Translational Research Center, Keio University School of Medicine, Tokyo, Japan.|Kanagawa Academy of Science and Technology, Kanagawa, Japan.|Division of Hematology, Keio University School of Medicine, Tokyo, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Ono-Uruga
authors.affiliation: Division of Hematology, Keio University School of Medicine, Tokyo, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Tozawa
authors.affiliation: Clinical and Translational Research Center, Keio University School of Medicine, Tokyo, Japan.|Kanagawa Academy of Science and Technology, Kanagawa, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Horiuchi
authors.affiliation: Department of Laboratory Medicine, Keio University School of Medicine, Tokyo, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Murata
authors.affiliation: Division of Hematology, Keio University School of Medicine, Tokyo, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Okamoto
authors.affiliation: Division of Hematology, Keio University School of Medicine, Tokyo, Japan.|Faculty of Science and Engineering, Life Science and Medical Bioscience, Waseda University, Tokyo, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Ikeda
authors.affiliation: International Research Center for Medical Sciences, Kumamoto University, Kumamoto, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Suda
authors.affiliation: Clinical and Translational Research Center, Keio University School of Medicine, Tokyo, Japan.|Kanagawa Academy of Science and Technology, Kanagawa, Japan.|Department of Laboratory Medicine, Keio University School of Medicine, Tokyo, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Matsubara
mesh_terms: D000111:Acetylcysteine / Q000378:metabolism; D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001792:Blood Platelets / Q000166:cytology*; D001803:Blood Transfusion; D002454:Cell Differentiation; D019070:Cell Lineage; D018014:Gene Transfer Techniques; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology*; D051379:Mice; D034741:RNA, Small Interfering / Q000378:metabolism; D011990:Receptors, Transferrin / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology; D017154:Stromal Cells / Q000166:cytology*; D034061:Thrombopoiesis; D013926:Thrombopoietin / Q000378:metabolism*; D014162:Transfection; D014168:Transferrin / Q000378:metabolism
chemical_list: D015703:Antigens, CD; C068126:CD71 antigen; D034741:RNA, Small Interfering; D011990:Receptors, Transferrin; D014168:Transferrin; D013926:Thrombopoietin; D000111:Acetylcysteine
keywords: adipose tissue; megakaryocytes; platelets; thrombopoietin; transferrin