Introduction

Stem cell researchers try to mimic physiological conditions in vitro in order to expand our knowledge about the regenerative properties of stem cells. One physiological condition that can be replicated is the hypoxic conditions that can be found in tissue. The effects of low oxygen tension are species- and tissue source-dependent. For example, although low oxygen tension has been shown not to affect human mesenchymal stem cells (MSCs) phenotype¹, it has been reported that it influences proliferation kinetics and metabolism². With mouse MSCs, on the other hand, low oxygen tension has been shown to affect both the cell surface marker expression rate and expansion capability of mouse MSCs². These findings show that mimicking physiological conditions is key.

For inducing low oxygen tension, STEMCELL offers the (Catalog #27310).

Hypoxia Chamber Set-up



A. Gas Cylinder (external vendor)

You will need certified Medical Grade pre-mixed gas. For culturing in 5% oxygen, we recommend ordering a nitrogen tank, with the following hypoxic mixture: 5% O₂, 10% CO₂, 85% N₂. If a lower oxygen percentage is required, for example 2% oxygen instead of 5%, then adjust the N₂% to make up the balance. So in this case, increase the N₂ to 88% from 85%.


B. Tygon Tubing (external vendor)

You will need 5/16" interior diameter Tygon tubing. This will be used between the gas tank and the single flow meter.


C. - optional

You may wish to use a single flow meter, which will allow you to measure the rate of gas going into the chamber. Attach the flow meter’s filter (included) to inlet flow meter tubing and then to gas tank. Adjust both the flow meter and regulator on the gas tank to allow a flow rate of 20-25 liters/min. (Maximum flow rate is 25 LPM). Connect gas outlet tube (top tube) of the flow meter to the Hypoxia Incubator Chamber. Flush the system for four minutes. Disconnect the incubator from the flow meter. Seal the incubator by closing plastic clamps (white) of gas inlet and outlet ports on the incubator.


D. (Catalog #27310)

The hypoxia incubator chamber is designed to maintain a hypoxic environment for cell culture. The chamber holds up to 12 cell culture plates.

After flushing the Chamber with the desired gas mixture, the Chamber seals tightly due to the O-ring and clamp, and your cultures are maintained in low oxygen conditions. Please see this instructional video for help assembling, flushing and sealing the Chamber, .