Orderly arrangement of hepatocyte spheroids on a microfabricated chip
Orderly arrangement of hepatocyte spheroids on a microfabricated chip
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DOI：
10.1089/ten.2005.11.1254
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发表时间：
2005-07-01
影响因子：
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通讯作者：
Nakazawa, K
中科院分区：
文献类型：
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作者：
Fukuda, J;Nakazawa, K
本文介绍了一种新的方法，用于制备几个球形多细胞聚集体（球体），具有几乎相同的直径上的微加工芯片。该芯片，通过一个简单的方法，使用微铣削系统制造，包括几千个空腔，直径100 - 500 μ m，在一个三角形排列的聚苯乙烯板。虽然在静止条件下培养时在任何芯片上均未形成球状体，但当用旋转振荡器施加旋转力时，肝细胞在空腔中形成球状体。特别是在具有直径为300 μ m的空腔的芯片上，每个空腔中形成一个球状体;构造了几乎相同直径的球状体的有序阵列（1100个球状体/ cm（2））。芯片上球状体的氨去除和白蛋白分泌在培养的至少14天内继续以初始水平发生。球状体的酶P- 450活性也得以保持，并在该透明芯片上通过由乙氧基试卤灵转化的试卤灵的荧光进行检测。该微阵列芯片是一种很有前途的细胞平台，可用于各种生物医学应用，如用于毒理学和药理学检查的基于细胞的生物传感器，以及生物人工肝。
This article describes a novel method for preparing several spherical multicellular aggregates ( spheroids) that have almost the same diameter on a microfabricated chip. The chip, fabricated by a simple method that uses a micromilling system, consisted of several thousand cavities, 100 - 500 mu m in diameter, in a triangular arrangement on a polystyrene plate. Although no spheroid was formed on any chip when cultured under stationary conditions, hepatocytes formed spheroids in the cavities when turning force was applied with a rotary shaker. Especially on the chip with cavities 300 mu m in diameter, one spheroid formed in each cavity; an orderly array of spheroids ( 1100 spheroids/ cm(2)) of almost the same diameter was constructed. Ammonia removal and albumin secretion by the spheroids on the chip continued to occur at initial levels for at least 14 days of culture. Enzyme P- 450 activity of the spheroids was also maintained and detected on this transparent chip by fluorescence of resorufin converted from ethoxyresorufin. The spheroid microarray chip seems to be a promising cellular platform for various biomedical applications such as in cell- based biosensors for toxicological and pharmacological examinations, and in bioartificial livers.