Layer-by-layer Collagen Deposition in Microfluidic Devices for Microtissue Stabilization

William J. McCarty; Ljupcho Prodanov; Shyam Sundhar Bale; Abhinav Bhushan; Rohit Jindal; Martin L. Yarmush; O. Berk Usta

doi:10.3791/53078

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

Layer-by-layer Collagen Deposition in Microfluidic Devices for Microtissue Stabilization

层与层胶原沉积在微流体装置的Microtissue稳定

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09:56 min

September 29, 2015

DOI: 10.3791/53078-v

William J. McCarty^1,2, Ljupcho Prodanov^1,2, Shyam Sundhar Bale^1,2, Abhinav Bhushan^1,2, Rohit Jindal^1,2, Martin L. Yarmush^1,2, O. Berk Usta^1,2

¹Center for Engineering in Medicine,Massachusetts General Hospital, Harvard Medical School, ²Shriners Hospitals for Children-Boston

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在微流控器件中创造功能性微组织需要通过将传统细胞培养技术适应微器件中有限的空间维度来稳定细胞表型。胶原蛋白的修饰允许超薄胶原蛋白组装体逐层沉积，这些组装体可以稳定原代细胞，例如肝细胞，作为微流体组织模型。

该程序的总体目标是将薄的胶原蛋白组件沉积到微流体装置中的细胞上。这是通过首先通过甲基化修饰酸化的天然胶原蛋白以产生带正电的净胶原蛋白分子来实现的。第二步是通过修饰酸化的天然胶原蛋白，以产生带负电的胶原蛋白分子。

接下来，制备微流体装置并接种肝细胞，使其附着并扩散过夜。最后一步是通过交替将细胞暴露于带正电和带负电的胶原蛋白溶液来沉积胶原蛋白组件，从而在细胞层顶部产生 10 个双层胶原蛋白。最终，相和免疫荧光显微镜以及白蛋白和尿素测定用于显示细胞极性和分泌功能的发育和维持。

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生物工程第103 胶原蛋白层 - 层沉积微流体肝细胞甲基化琥珀酰化肝microtissue