Dynamic single cell culture array - PubMed (original) (raw)

doi: 10.1039/b605937f. Epub 2006 Sep 4.

Affiliations

PMID: 17066168
DOI: 10.1039/b605937f

Dynamic single cell culture array

Dino Di Carlo et al. Lab Chip. 2006 Nov.

Abstract

It is important to quantify the distribution of behavior amongst a population of individual cells to reach a more complete quantitative understanding of cellular processes. Improved high-throughput analysis of single cell behavior requires uniform conditions for individual cells with controllable cell-cell interactions, including diffusible and contact elements. Uniform cell arrays for static culture of adherent cells have previously been constructed using protein micropatterning techniques but lack the ability to control diffusible secretions. Here we present a microfluidic-based dynamic single cell culture array that allows both arrayed culture of individual adherent cells and dynamic control of fluid perfusion with uniform environments for individual cells. In our device no surface modification is required and cell loading is done in less than 30 seconds. The device consists of arrays of physical U-shaped hydrodynamic trapping structures with geometries that are biased to trap only single cells. HeLa cells were shown to adhere at a similar rate in the trapping array as on a control glass substrate. Additionally, rates of cell death and division were comparable to the control experiment. Approximately 100 individual isolated cells were observed growing and adhering in a field of view spanning approximately 1 mm(2) with greater than 85% of cells maintained within the primary trapping site after 24 hours. Also, greater than 90% of cells were adherent and only 5% had undergone apoptosis after 24 hours of perfusion culture within the trapping array. We anticipate uses in single cell analysis of drug toxicity with physiologically relevant perfused dosages as well as investigation of cell signaling pathways and systems biology.

PubMed Disclaimer

Cited by

Multiparameter cell affinity chromatography: separation and analysis in a single microfluidic channel.
Li P, Gao Y, Pappas D. Li P, et al. Anal Chem. 2012 Oct 2;84(19):8140-8. doi: 10.1021/ac302002a. Epub 2012 Sep 21. Anal Chem. 2012. PMID: 22958145 Free PMC article.
Real-time analysis and selection of methylated DNA by fluorescence-activated single molecule sorting in a nanofluidic channel.
Cipriany BR, Murphy PJ, Hagarman JA, Cerf A, Latulippe D, Levy SL, Benítez JJ, Tan CP, Topolancik J, Soloway PD, Craighead HG. Cipriany BR, et al. Proc Natl Acad Sci U S A. 2012 May 29;109(22):8477-82. doi: 10.1073/pnas.1117549109. Epub 2012 May 14. Proc Natl Acad Sci U S A. 2012. PMID: 22586076 Free PMC article.
Dual transcript and protein quantification in a massive single cell array.
Park SM, Lee JY, Hong S, Lee SH, Dimov IK, Lee H, Suh S, Pan Q, Li K, Wu AM, Mumenthaler SM, Mallick P, Lee LP. Park SM, et al. Lab Chip. 2016 Oct 7;16(19):3682-8. doi: 10.1039/c6lc00762g. Epub 2016 Aug 22. Lab Chip. 2016. PMID: 27546183 Free PMC article.
On-chip immunofluorescence analysis of single cervical cells using an electroactive microwell array with barrier for cervical screening.
Takeuchi M, Nagasaka K, Yoshida M, Kawata Y, Miyagawa Y, Tago S, Hiraike H, Wada-Hiraike O, Oda K, Osuga Y, Fujii T, Ayabe T, Kim SH, Fujii T. Takeuchi M, et al. Biomicrofluidics. 2019 Jul 30;13(4):044107. doi: 10.1063/1.5089796. eCollection 2019 Jul. Biomicrofluidics. 2019. PMID: 31431817 Free PMC article.
Optimization of Mechanical Tissue Dissociation Using an Integrated Microfluidic Device for Improved Generation of Single Cells Following Digestion.
Aliaghaei M, Haun JB. Aliaghaei M, et al. Front Bioeng Biotechnol. 2022 Feb 8;10:841046. doi: 10.3389/fbioe.2022.841046. eCollection 2022. Front Bioeng Biotechnol. 2022. PMID: 35211466 Free PMC article.

Dynamic single cell culture array - PubMed (original) (raw)