Design and Assessment of a Dynamic Perfusion Bioreactor for Large Bone Tissue Engineering Scaffolds
Design and Assessment of a Dynamic Perfusion Bioreactor for Large Bone Tissue Engineering Scaffolds
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DOI：
10.1007/s12010-017-2671-5
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发表时间：
2018-06-01
影响因子：
3
通讯作者：
Reilly, Gwendolen C.
中科院分区：
文献类型：
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作者：
Bhaskar, Birru;Owen, Robert;Reilly, Gwendolen C.
生物反应器可用于在体外将流体流施加到支架，以改善培养基的质量输送并向细胞施加机械力。在这项研究中，我们开发并测试了一种适用于大型支架的可高压灭菌的模块化灌注生物反应器。我们研究了流体流动诱导的剪切应力（FFSS）对人胚胎干细胞衍生的间充质祖细胞（hES-MP细胞）在成骨诱导培养基（OIM）中的大聚氨酯（PU）支架（30 mm直径× 5 mm厚度）上培养的成骨分化的影响。接种后，将支架保持在静态条件下或在接种后3天转移到生物反应器中，并应用3.47 mL/min的连续流速。碱性磷酸酶活性（ALP）用于评价成骨分化和刃天青盐减少（RR），以测量10天后的代谢活性。与静态培养中生长的支架相比，流动培养物含有更多的代谢活性细胞和更高的总DNA含量，以及更高的ALP活性。这些结果证实了hES-MP细胞对流体流动刺激的响应性，并提出了一种具有成本效益的、用户友好的生物反应器，该生物反应器能够支持间充质祖细胞在能够填充大骨缺损的支架内的生长和分化。
Bioreactors can be used to apply fluid flow in vitro to scaffolds to improve mass transport of media and apply mechanical forces to cells. In this study, we developed and tested an autoclavable, modular perfusion bioreactor suitable for large scaffolds. We investigated the effects of fluid flow induced shear stress (FFSS) on osteogenic differentiation of human embryonic stem cell-derived mesenchymal progenitors (hES-MP cells) cultured on large polyurethane (PU) scaffolds (30 mm diameter x 5 mm thickness) in osteogenesis induction media (OIM). After seeding, scaffolds were either maintained in static conditions or transferred to the bioreactor 3 days post-seeding and a continuous flow rate of 3.47 mL/min was applied. Alkaline phosphatase activity (ALP) was used to evaluate osteogenic differentiation and resazurin salt reduction (RR) to measure metabolic activity after 10 days. Cultures subjected to flow contained significantly more metabolically active cells and higher total DNA content, as well as significantly higher ALP activity compared to scaffolds grown in static culture. These results confirm the responsiveness of hES-MP cells to fluid flow stimuli, and present a cost-effective, user-friendly bioreactor capable of supporting the growth and differentiation of mesenchymal progenitor cells within scaffolds capable of filling large bone defects.