Low-cost, open-source cell culture chamber for regulating physiologic oxygen levels

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 O 2 between 1 and 21%, while maintaining CO 2 levels at 5%, as well as monitoring of temperature, pressure, and relative humidity. Testing showed our hypoxia chamber was able to maintain CO 2 levels at 5% and hypoxic O 2 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).

A modular and flexible open source cell incubator system for mobile and stationary use

“…Some of these incubators focus on achieving lower oxygen concentrations than what is present in ambient air, as such conditions can aid cell growth and viability under certain conditions [19]. These conditions are more physiological and can either be created passively by mixing O 2 and N 2 in an open-loop fashion [20], or by actively controlling the corresponding gas concentrations [21]. The latter work has also suggested to flexibly adapt the incubation chamber.…”

Robotic System for Organoid Assembly in a Multi-Well Microfluidic Chip

“…While commercial miniature CO2 incubators exist that could fit in a hood, such as the MyTemp Mini CO2 Digital Incubator (Benchmark Scientific), these are not structurally modifiable, and therefore not appropriate for a prototyping system that might need to expand in size and be controlled in unconventional ways. Recently, several labs have constructed DIY CO2 incubators, [24][25][26][27][28] and the Pelling Lab implementation led to the company Incuvers that produces modifiable Arduinocontrolled incubators, 29 from which we acquired the Incuvers Model 1 (Figure 2A).…”

Development of a Smart Portable Hypoxic Chamber with Accurate Sensing, Control and Visualization of In Vitro Cell Culture for Replication of Cancer Microenvironment

“…A challenge thus arises in replicating and maintaining the hypoxic conditions when testing drugs for these diseases, which now becomes a vital component in cancer models and treatment studies [19]. To induce such conditions, hypoxic chambers are used as an alternative to chemical mimetic agents [20,21]. These are the devices that help maintain a specified amount of oxygen condition without compromising cell viability.…”

“…The issue with these devices, however, is that there is no inbuilt method to measure or monitor the oxygen levels inside the chamber during the entirety of the experiment, and a need for cost effective and controlled hypoxia chambers then arises. Various research has been conducted to measure and record the oxygen percentage in the chamber during the experiment duration [20,[22][23][24]. Though this helps in monitoring the conditions of experiment, it still lacks an aspect of control.…”