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  • Ultrasonic three-dimensional on-chip cell culture for dynamic studies of tumor immune surveillance by natural killer cells.

Ultrasonic three-dimensional on-chip cell culture for dynamic studies of tumor immune surveillance by natural killer cells.

Lab on a chip (2015-07-02)
Athanasia E Christakou, Mathias Ohlin, Björn Önfelt, Martin Wiklund
摘要

We demonstrate a simple method for three-dimensional (3D) cell culture controlled by ultrasonic standing waves in a multi-well microplate. The method gently arranges cells in a suspension into a single aggregate in each well of the microplate and, by this, nucleates 3D tissue-like cell growth for culture times between two and seven days. The microplate device is compatible with both high-resolution optical microscopy and maintenance in a standard cell incubator. The result is a scaffold- and coating-free method for 3D cell culture that can be used for controlling the cellular architecture, as well as the cellular and molecular composition of the microenvironment in and around the formed cell structures. We demonstrate the parallel production of one hundred synthetic 3D solid tumors comprising up to thousands of human hepatocellular carcinoma (HCC) HepG2 cells, we characterize the tumor structure by high-resolution optical microscopy, and we monitor the functional behavior of natural killer (NK) cells migrating, docking and interacting with the tumor model during culture. Our results show that the method can be used for determining the collective ability of a given number of NK cells to defeat a solid tumor having a certain size, shape and composition. The ultrasound-based method itself is generic and can meet any demand from applications where it is advantageous to monitor cell culture from production to analysis of 3D tissue or tumor models using microscopy in one single microplate device.

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Sigma-Aldrich
青链霉素, Solution stabilized, with 10,000 units penicillin and 10 mg streptomycin/mL, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
L -谷氨酰胺 溶液, 200 mM, solution, sterile-filtered, BioXtra, suitable for cell culture
SAFC
MEM 非必需氨基酸溶液 (100×), without L-glutamine, liquid, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
丙酮酸钠 溶液, 100 mM, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
丙酮酸钠, ReagentPlus®, ≥99%
Sigma-Aldrich
Iscove 改良杜氏培养基, liquid, sterile-filtered, With sodium bicarbonate, without L-glutamine, suitable for cell culture, suitable for hybridoma
Sigma-Aldrich
L-谷氨酰胺, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
Sigma-Aldrich
丙酮酸钠, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99%
Sigma-Aldrich
L-谷氨酰胺, ReagentPlus®, ≥99% (HPLC)
Sigma-Aldrich
钙黄绿素, Used for the fluorometric determination of calcium and EDTA titration of calcium in the presence of magnesium.
SAFC
L-谷氨酰胺
Sigma-Aldrich
丙酮酸钠, powder, BioXtra, suitable for mouse embryo cell culture
Sigma-Aldrich
L-谷氨酰胺, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
丙酮酸钠, BioXtra, ≥99%
Sigma-Aldrich
L-谷氨酰胺, γ-irradiated, BioXtra, suitable for cell culture
Sigma-Aldrich
丙酮酸钠, Hybri-Max, powder, suitable for hybridoma
Sigma-Aldrich
L-谷氨酰胺
Sigma-Aldrich
丙酮酸钠, anhydrous, free-flowing, Redi-Dri, ReagentPlus®, ≥99%