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一种用于三维细胞培养和基于细胞的分析的微流控平台。

A microfluidic platform for 3-dimensional cell culture and cell-based assays.

作者信息

Kim Minseok S, Yeon Ju Hun, Park Je-Kyun

机构信息

Department of BioSystems, Korea Advanced Institute of Science and Technology (KAIST), 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea.

出版信息

Biomed Microdevices. 2007 Feb;9(1):25-34. doi: 10.1007/s10544-006-9016-4.

Abstract

This paper reports a novel microfluidic platform introducing peptide hydrogel to make biocompatible microenvironment as well as realizing in situ cell-based assays. Collagen composite, OPLA and Puramatrix scaffolds are compared to select good environment for human hepatocellular carcinoma cells (HepG2) by albumin measurement. The selected biocompatible self-assembling peptide hydrogel, Puramatrix, is hydrodynamically focused in the middle of main channel of a microfluidic device, and at the same time the cells are 3-dimensionally immobilized and encapsulated without any additional surface treatment. HepG2 cells have been 3-dimensionally cultured in a poly(dimethylsiloxane) (PDMS) microfluidic device for 4 days. The cells cultured in micro peptide scaffold are compared with those cultured by conventional petri dish in morphology and the rate of albumin secretion. By injection of different reagents into either side of the peptide scaffold, the microfluidic device also forms a linear concentration gradient profile across the peptide scaffold due to molecular diffusion. Based on this characteristic, toxicity tests are performed by Triton X-100. As the higher toxicant concentration gradient forms, the wider dead zone of cells in the peptide scaffold represents. This microfluidic platform facilitates in vivo-like 3-dimensional microenvironment, and have a potential for the applications of reliable cell-based screening and assays including cytotoxicity test, real-time cell viability monitoring, and continuous dose-response assay.

摘要

本文报道了一种新型微流控平台,该平台引入肽水凝胶以形成生物相容性微环境,并实现基于细胞的原位分析。通过测量白蛋白,比较了胶原蛋白复合材料、OPLA和Puramatrix支架,以选择适合人肝癌细胞(HepG2)生长的良好环境。所选择的具有生物相容性的自组装肽水凝胶Puramatrix在微流控装置主通道的中间实现流体动力学聚焦,同时细胞在无需任何额外表面处理的情况下被三维固定和包封。HepG2细胞已在聚二甲基硅氧烷(PDMS)微流控装置中进行了4天的三维培养。将在微肽支架中培养的细胞与在传统培养皿中培养的细胞在形态和白蛋白分泌率方面进行了比较。通过向肽支架的两侧注入不同的试剂,由于分子扩散,微流控装置还在肽支架上形成了线性浓度梯度分布。基于这一特性,使用Triton X-100进行了毒性测试。随着更高的毒物浓度梯度形成,肽支架中细胞的死亡区域越宽。这种微流控平台有助于形成类似体内的三维微环境,并具有在基于细胞的可靠筛选和分析中的应用潜力,包括细胞毒性测试、实时细胞活力监测和连续剂量反应分析。

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