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首页> 外文期刊>Biomaterials >A 3D microfibrous scaffold for long-term human pluripotent stem cell self-renewal under chemically defined conditions
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A 3D microfibrous scaffold for long-term human pluripotent stem cell self-renewal under chemically defined conditions

机译：3D微纤维支架，可在化学定义的条件下使人类多能干细胞长期自我更新

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摘要

Realizing the potential of human pluripotent stem cell (hPSC)-based therapy requires the development of defined scalable culture systems with efficient expansion, differentiation and isolation protocols. We report an engineered 3D microfiber system that efficiently supports long-term hPSCs self-renewal under chemically defined conditions. The unique feature of this system lies in the application of a 3D ECM-like environment in which cells are embedded, that affords: (i) uniform high cell loading density in individual cell-laden constructs (～10 7 cells/ml); (ii) quick recovery of encapsulated cells (10min at 37°C) with excellent preservation of cell viability and 3D multicellular structure; (iii) direct cryopreservation of the encapsulated cells in situ in the microfibers with 17-fold higher cell viability compared to those cultured on Matrigel surface; (iv) long-term hPSC propagation under chemically defined conditions. Four hPSC lines propagated in the microfibrous scaffold for 10 consecutive passages were capable of maintaining an undifferentiated phenotype as demonstrated by the expression of stem cell markers and stable karyotype invitro and the ability to form derivatives of the three germ layers both invitro and invivo. Our 3D microfibrous system has the potential for large-scale cultivation of transplantable hESCs and derivatives for clinical applications.
机译：要实现基于人多能干细胞（hPSC）疗法的潜力，就需要开发定义好的可扩展培养系统，并具有有效的扩增，分化和分离方案。我们报告了一种经过工程设计的3D超细纤维系统，该系统可在化学定义的条件下有效支持hPSC的长期自我更新。该系统的独特之处在于可在3D ECM样环境中嵌入细胞，该环境可提供：（i）各个载有细胞的构建体中均一的高细胞装载密度（约10 7个细胞/ ml）; （ii）快速回收被包封的细胞（在37°C下<10分钟），并具有出色的细胞活力和3D多细胞结构保存能力; （iii）与在Matrigel表面培养的细胞相比，将被包裹的细胞直接冷冻保存在微纤维中，其细胞活力高出17倍以上; （iv）hPSC在化学条件下的长期繁殖。在微纤维支架中繁殖10个连续传代的4个hPSC系能够维持未分化的表型，如干细胞标记物的表达和稳定的核型在体外以及在体外和体内形成三个胚层的衍生物的能力所证明。我们的3D微纤维系统具有用于临床应用的可移植hESC及其衍生物大规模培养的潜力。

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