Abstract
The physiological oxygen levels for several mammalian cell types in vivo are considered to be hypoxic (low oxygen tension), but the vast majority of in vitro mammalian cell culture is conducted at atmospheric oxygen levels of around 21%. In order to understand the impact of low oxygen environments on cells, oxygen levels need to be regulated during in vitro culture. Two common methods for simulating a hypoxic environment are through the regulation of gas composition or chemical induction. Chemically mimicking hypoxia can have adverse effects such as reducing cell viability, making oxygen regulation in cell culture chambers crucial for long-term culture. However, oxygen-regulating cell culture incubators and commercial hypoxia chambers may not always be a viable option due to cost and limited customization. Other low-cost chambers have been developed, but they tend to lack control systems or are fairly small scale. Thus, the objective of this project was to design and develop a low-cost, open-source, controllable, and reproducible hypoxia chamber that can fit inside a standard cell culture incubator. This design allows for the control of O2 between 1 and 21%, while maintaining CO2 levels at 5%, as well as monitoring of temperature, pressure, and relative humidity. Testing showed our hypoxia chamber was able to maintain CO2 levels at 5% and hypoxic O2 levels at 1% and 5% for long-term cell culture. This simple and easy-to-manufacture design uses off the shelf components, and the total material cost was $832.47 (USD).
Highlights
The physiological oxygen levels for several mammalian cell types in vivo are considered to be hypoxic, but the vast majority of in vitro mammalian cell culture is conducted at atmospheric oxygen levels of around 21%
Other low-cost chambers have been developed, but they tend to lack control systems or are fairly small scale. The objective of this project was to design and develop a low-cost, open-source, controllable, and reproducible hypoxia chamber that can fit inside a standard cell culture incubator
Induced hypoxia aims to mimic the response of hypoxia inducible factor (HIF)-1, which is a transcription factor activated in cells by hypoxic environments [34]
Summary
The design files include three-dimensional (3D) drawings of all components as STL and AutodeskÒ Fusion 360 files (F3Z and F3D), as well as printed circuit board (PCB) design files, code for the controllers and GUI as outlined and described below and in the Design Files Summary. It includes an Arduino Mega, the O2 and CO2 sensors, a logic level converter to ensure the 3.3 V UART signals of the sensors can communicate with the 5 V Arduino signals, three solenoids, 1kO resistor, BJT transistor, and a diode for each of the solenoids. The Arduino has two files written in the default Arduino IDE that get compiled together to run and control the Arduino. The Raspberry Pi has a python file that runs the GUI. GUI.py was created using the tkinter library in Python
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