高通量基于 3D 细胞的增殖和细胞毒性测定在药物筛选和生物工艺开发中的应用。

High-throughput 3-D cell-based proliferation and cytotoxicity assays for drug screening and bioprocess development.

机构信息

William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 West 19th Avenue, Columbus, OH 43210, USA.

出版信息

J Biotechnol. 2011 Jan 20;151(2):186-93. doi: 10.1016/j.jbiotec.2010.11.012. Epub 2010 Nov 27.

DOI:10.1016/j.jbiotec.2010.11.012

PMID:21115074

Abstract

We have designed, built and tested a three-dimensional (3-D) cell culture system on modified microplates for high-throughput, real-time, proliferation and cytotoxicity assays. In this 3-D culture system, cells expressing the enhanced green fluorescent protein (EGFP) were cultured in nonwoven polyethylene terephthalate (PET) fibrous scaffolds. Compared to 2-D cultures in conventional microplates, 3-D cultures gave more than 10-fold higher fluorescence signals with significantly increased signal-to-noise ratio (SNR), thus extending the application of conventional fluorescence microplate readers for online monitoring of culture fluorescence. The 3-D system was successfully used to demonstrate the effects of fetal bovine serum, fibronectin coating of PET fibers, and cytotoxicity of dexamethasone on recombinant murine embryonic stem D3 cells. The dosage effects of 5-fluorouracil and gemcitabine on high-density colon cancer HT-29 cells were also tested. These studies demonstrated that the 3-D culture microplate system with EGFP expressing cells can be used as a high-throughput system in drug discovery and bioprocess development.

摘要

我们设计、构建并测试了一种基于改良微板的三维（3-D）细胞培养系统，用于高通量、实时的增殖和细胞毒性检测。在这个 3-D 培养系统中，表达增强型绿色荧光蛋白（EGFP）的细胞在无纺聚酯纤维（PET）纤维支架中培养。与传统微板中的 2-D 培养相比，3-D 培养产生的荧光信号高出 10 倍以上，且信号与噪声比（SNR）显著增加，从而扩展了传统荧光微板读数器在培养荧光在线监测方面的应用。该 3-D 系统成功地用于展示胎牛血清、PET 纤维的纤连蛋白涂层以及地塞米松对重组鼠胚胎干细胞 D3 细胞的细胞毒性的影响。还测试了 5-氟尿嘧啶和吉西他滨对高密度结肠癌 HT-29 细胞的剂量效应。这些研究表明，具有 EGFP 表达细胞的 3-D 培养微板系统可用作药物发现和生物工艺开发的高通量系统。