Host: The Japanese Society of Toxicology
Name : The 49th Annual Meeting of the Japanese Society of Toxicology
Date : June 30, 2022 - July 02, 2022
1.Introduction
Three-dimensional liver spheroids are expected to be nearly a living body in terms of drug responsiveness compared to planar culture.
2.Objective
In this study, we aimed to evaluate the drug response by a liver spheroid with a new pore structure in which hydrogel beads were dissolved, expecting a higher efficiency of mass exchange inside the spheroid.
3.Method
Spheroids were produced by dispensing 1 µl of HuH-7 cell, a human hepatoma cell line, suspending normal medium into 3% methylcellulose medium. [ref 1]
And, the hydrogel beads embedding in the hybrid spheroids were dissolved with alginate lyase to produce spheroids with a new pore structure.
After one day of incubation in methylcellulose, spheroids were removed by cellulase treatment and treated with chemicals.
Mitomycin C was utilized as drug, and their concentrations in the assay was 10μM and 100 μM, respectively.
The living and dead cells in the spheroids were visualized by LIVE/DEAD assay in combination with confocal laser microscopy and measured by image analysis.
4.Results
The cell death rate of mitomycin C was about 10% higher in spheroids with pore structure than in spheroids without pore structure.
The cell death rates of spheroids with hydrogel beads and with a new pore structure by dissolving hydrogel beads were almost the same and not significantly different.
[ref 1] Kojima, N., Takeuchi, S. and Sakai, Y. Rapid aggregation of heterogeneous cells and multiple-sized microspheres in methylcellulose medium. Biomaterials, 33, 4508-4514 (2012).
