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Issue 9, 2018, Issue in Progress
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Pitch-tunable pillar arrays for high-throughput culture and immunohistological analysis of tumor spheroids

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Abstract

Tumor spheroids are multicellular, three-dimensional (3D) cell culture models closely mimicking the microenvironments of human tumors in vivo, thereby providing enhanced predictability, clinical relevancy of drug efficacy and the mechanism of action. Conventional confocal microscopic imaging remains inappropriate for immunohistological analysis due to current technical limits in immunostaining using antibodies and imaging cells grown in 3D multicellular contexts. Preparation of microsections of these spheroids represents a best alternative, yet their sub-millimeter size and fragility make it less practical for high-throughput screening. To address these problems, we developed a pitch-tunable 5 × 5 mini-pillar array chip for culturing and sectioning tumor spheroids in a high throughput manner. Tumor spheroids were 3D cultured in an alginate matrix using a twenty-five mini-pillar array which aligns to a 96-well. At least a few tens of spheroids per pillar were cultured and as many as 25 different treatment conditions per chip were evaluated, which indicated the high throughput manner of the 5 × 5 pillar array chip. The twenty-five mini-pillars were then rearranged to a transferring pitch so that spheroid-containing gel caps from all pillars can be embedded into a specimen block. Tissue array sections were then prepared and stained for immunohistological examination. The utility of this pitch-tunable pillar array was demonstrated by evaluating drug distribution and expression levels of several proteins following drug treatment in 3D tumor spheroids. Overall, our mini-pillar array provides a novel platform that can be useful for culturing tumor spheroids as well as for immunohistological analysis in a multiplexed and high throughput manner.

Graphical abstract: Pitch-tunable pillar arrays for high-throughput culture and immunohistological analysis of tumor spheroids

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Supplementary files

Article information


Submitted
17 Aug 2017
Accepted
08 Jan 2018
First published
24 Jan 2018

This article is Open Access

RSC Adv., 2018,8, 4494-4502
Article type
Paper

Pitch-tunable pillar arrays for high-throughput culture and immunohistological analysis of tumor spheroids

D. W. Lee, J. Kang, H. J. Hwang, M. Oh, B. C. Shin, M. Lee and H. Kuh, RSC Adv., 2018, 8, 4494
DOI: 10.1039/C7RA09090K

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    [Original citation] - Published by The Royal Society of Chemistry.

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