期刊
CELL CALCIUM
卷 47, 期 3, 页码 224-233出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.ceca.2009.11.012
关键词
High content screening; Calcium imaging; Cardiac myocytes; Multi-well plates; TIRE; Elastic coating
类别
资金
- Federal Ministry of Education and Research (BMBF, Germany)
- Federal Institute for Risk Assessment (BfR, Germany)
- German Research Foundation (DFG) [KFO 196]
- Collaborative Research Centre [SFB 530]
The application of primary excitable cells for high content screening (HCS) requires a multitude of novel developments including cell culture and multi-well plates. Here we introduce a novel system combining optimised culture conditions of primary adult cardiomyocytes with the particular needs of excitable cells for arbitrary field stimulation of individual wells. The major advancements of our design were tested in calcium imaging experiments and comprise (i) each well of the plate can be subjected to individual pulse protocols, (ii) the software driving electrical stimulation can run as a stand-alone application but also as a plug-into HCS software packages, (iii) the optical properties of the plastic substrate (foil) resemble those of glass coverslips fostering high resolution immersion-based microscopy, (iv) the bottom of the foil is coated with an oleophobic layer that prevents immersion oil from sticking, (v) the top of the foil is coated with an elastic film. The latter enables cardiomyocytes to display loaded contractions by mimicking the physiologically occurring local elastic network (e.g. extracellular matrix) and results in significantly increased contractions (with identical calcium transients) when compared to non-elastic substrates. Thus, our novel design and culture conditions represent an essential further step towards the application of primary cultured adult cardiornyocytes for HCS applications. (C) 2009 Elsevier Ltd. All rights reserved.
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