Abstract
Purpose :
In the ophthalmological cell biology research, the oxygen concentration is one of the important parameters. The corneal epithelial cells in the human eye are exposed to the air with around 20% oxygen concentration. But, some cells could not be supported the sufficient oxygen because of a contact lens in front of their cells. To mimic the real environment at an in-vitro cell-based experiment, we developed the automatic real-time controlled hypoxia system based on a multi-well plate with diverse oxygen tension on each well.
Methods :
The present high throughput hypoxia screening(HTHS) system is applied with immortalized human corneal epithelial cell line (HCE-Ts) with reperfusion condition for 24 hours after exposing hypoxia condition with 2, 5, 10% oxygen concentrations each during another 24 hours. The other cells as control groups had been keeping hypoxia condition continuously during 48 hours on a 6-well plate at the same time. This HTHS system with 6 different oxygen concentration each is fully automatically operated by the programmed a microprocessor (Arduino MEGA) without any handling during the entire experimental process.
Results :
The high throughput hypoxia screening(HTHS) system for ophthalmological in-vitro cell study: Cytotoxicity by oxidative stress on HCE-Ts cells after hypoxia environment was shown with two times increase compared to control cells under keeping hypoxia conditions as a control group. We have been repeating these experiments with more diverse oxidative stress on conjunctival cells to mimic the real environment on in-vitro experiment.
Conclusions :
We have developed a high throughput hypoxia screening (HTHS) system for cytotoxicity screening of oxidative stress with 6 different oxygen concentrations. Our HTHS system can obtain more progressed oxidative stress results with minimizing the consumption of time, and labor. This system can be useful for hypoxia screening the new drug against diverse ophthalmological cell type instead of the human eye.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.