Megakaryocytes and platelets from a novel human adipose tissue–derived mesenchymal stem cell line

Keiichi Tozawa, Yukako Ono-Uruga, Masaki Yazawa, Taisuke Mori, Mitsuru Murata, Shinichiro Okamoto, Yasuo Ikeda, Yumiko Matsubara

研究成果: Article

7 引用 (Scopus)

抄録

The clinical need for platelet transfusions is increasing; however, donor-dependent platelet transfusions are associated with practical problems, such as the limited supply and the risk of infection. Thus, we developed a manufacturing system for platelets from a donor-independent cell source: a human adipose-derived mesenchymal stromal/stem cell line (ASCL). The ASCL was obtained using an upside-down culture flask method and satisfied the minimal criteria for defining mesenchymal stem cells (MSCs) by The International Society for Cellular Therapy. The ASCL showed its proliferation capacity for ‡2 months without any abnormal karyotypes. The ASCL was cultured in megakaryocyte induction media. ASCL-derived megakaryocytes were obtained, with a peak at day 8 of culture, and ASCL-derived platelets (ASCL-PLTs) were obtained, with a peak at day 12 of culture. We observed that CD42b 1 cells expressed an MSC marker (CD90) which is related to cell adhesion. Compared with peripheral platelets, ASCL-PLTs exhibit higher levels of PAC1 binding, P-selectin surface exposure, ristocetin-induced platelet aggregation, and ADP-induced platelet aggregation, as well as similar levels of fibrinogen binding and collagen-induced platelet aggregation. ASCL-PLTs have lower epinephrine-induced platelet aggregation. The pattern of in vivo kinetics after infusion into irradiated immunodeficient NOD.Cg-Prkdc scid Il2rg tm1Wjl / SzJ mice was similar to that of platelet concentrates. ASCL-PLTs have similar characteristics to those of peripheral platelets and might have an additional function as MSCs. The establishment of the ASCL and its differentiation into ASCL-PLTs do not require gene transfer, and endogenous thrombopoietin is used for differentiation. The present protocol is a simple method that does not require feeder cells, further enhancing the clinical application of our approach.

元の言語English
ページ(範囲)633-643
ページ数11
ジャーナルBlood
133
発行部数7
DOI
出版物ステータスPublished - 2019 2 14

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Megakaryocytes
Platelets
Stem cells
Mesenchymal Stromal Cells
Blood Platelets
Cell Line
Agglomeration
Platelet Aggregation
Cell culture
Platelet Transfusion
Ristocetin
Gene transfer
Feeder Cells
Thrombopoietin
P-Selectin
Cell adhesion
Adenosine Diphosphate
Fibrinogen
Epinephrine

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

これを引用

Megakaryocytes and platelets from a novel human adipose tissue–derived mesenchymal stem cell line. / Tozawa, Keiichi; Ono-Uruga, Yukako; Yazawa, Masaki; Mori, Taisuke; Murata, Mitsuru; Okamoto, Shinichiro; Ikeda, Yasuo; Matsubara, Yumiko.

:: Blood, 巻 133, 番号 7, 14.02.2019, p. 633-643.

研究成果: Article

Tozawa, Keiichi ; Ono-Uruga, Yukako ; Yazawa, Masaki ; Mori, Taisuke ; Murata, Mitsuru ; Okamoto, Shinichiro ; Ikeda, Yasuo ; Matsubara, Yumiko. / Megakaryocytes and platelets from a novel human adipose tissue–derived mesenchymal stem cell line. :: Blood. 2019 ; 巻 133, 番号 7. pp. 633-643.
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AU - Tozawa, Keiichi

AU - Ono-Uruga, Yukako

AU - Yazawa, Masaki

AU - Mori, Taisuke

AU - Murata, Mitsuru

AU - Okamoto, Shinichiro

AU - Ikeda, Yasuo

AU - Matsubara, Yumiko

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AB - The clinical need for platelet transfusions is increasing; however, donor-dependent platelet transfusions are associated with practical problems, such as the limited supply and the risk of infection. Thus, we developed a manufacturing system for platelets from a donor-independent cell source: a human adipose-derived mesenchymal stromal/stem cell line (ASCL). The ASCL was obtained using an upside-down culture flask method and satisfied the minimal criteria for defining mesenchymal stem cells (MSCs) by The International Society for Cellular Therapy. The ASCL showed its proliferation capacity for ‡2 months without any abnormal karyotypes. The ASCL was cultured in megakaryocyte induction media. ASCL-derived megakaryocytes were obtained, with a peak at day 8 of culture, and ASCL-derived platelets (ASCL-PLTs) were obtained, with a peak at day 12 of culture. We observed that CD42b 1 cells expressed an MSC marker (CD90) which is related to cell adhesion. Compared with peripheral platelets, ASCL-PLTs exhibit higher levels of PAC1 binding, P-selectin surface exposure, ristocetin-induced platelet aggregation, and ADP-induced platelet aggregation, as well as similar levels of fibrinogen binding and collagen-induced platelet aggregation. ASCL-PLTs have lower epinephrine-induced platelet aggregation. The pattern of in vivo kinetics after infusion into irradiated immunodeficient NOD.Cg-Prkdc scid Il2rg tm1Wjl / SzJ mice was similar to that of platelet concentrates. ASCL-PLTs have similar characteristics to those of peripheral platelets and might have an additional function as MSCs. The establishment of the ASCL and its differentiation into ASCL-PLTs do not require gene transfer, and endogenous thrombopoietin is used for differentiation. The present protocol is a simple method that does not require feeder cells, further enhancing the clinical application of our approach.

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