High-density hiPSCs expansion supported by growth factors accumulation in a simple dialysis-culture platform
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- Fuad Torizal
- Department of Bioengineering and Department of Chemical Systems Engineering, The University of Tokyo
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- Qiao You Lau
- Department of Bioengineering, School of Engineering, The University of Tokyo
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- Masato Ibuki
- Regenerative Medicine and Cell Therapy Laboratories, Kaneka Corporation
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- Yoshikazu Kawai
- Regenerative Medicine and Cell Therapy Laboratories, Kaneka Corporation
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- Masato Horikawa
- Materials Research Laboratories, Nissan Chemical Corporation
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- Masataka Minami
- Materials Research Laboratories, Nissan Chemical Corporation
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- Ikki Horiguchi
- Department of Biotechnology, School of Engineering, Osaka University
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- Masaki Nishikawa
- Department of Chemical Systems Engineering, School of Engineering, The University of Tokyo
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- Yasuyuki Sakai
- University of Tokyo
説明
<jats:title>Abstract</jats:title> <jats:p>Three-dimensional aggregate-suspension culture can produce large numbers of human induced pluripotent stem cells (hiPSCs); however, use of expensive growth factors and method-induced mechanical stress potentially result in inefficient production costs and difficulties in preserving pluripotency. Here, we developed a simple, miniaturized, dual-compartment dialysis-culture device based on a conventional membrane-culture insert with deep well plates. The device allowed growth-factor accumulation and improved cell expansion up to ~ 32 × 10<jats:sup>6</jats:sup> cells/mL, and reduction of excessive shear stress and agglomeration following addition of the functional polymer FP003 supported high-density expansion. The results revealed accumulation of several growth factors, including fibroblast growth factor 2 and insulin, along with endogenous NODAL, which acts as a substitute for depleted transforming growth factor-β1 in maintaining pluripotency. Because we used the same growth-factor formulation per volume in the upper culture compartment, cost reduction increased significantly in proportional manner with cell density. We showed that growth-factor-accumulation dynamics in a low-shear-stress environment successfully improved hiPSC proliferation, pluripotency, and differentiation potential. This miniaturised dialysis-culture system demonstrated the feasibility and cost-effective mass production of hiPSCs in high-density culture.</jats:p>
