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Hypoxic Cell Culture System

Modular hypoxia equipment

In Vitro Hypoxia Considerations

Oxygen levels regulate many aspects of cellular function: metabolism, gene expression, cell cycle, proliferative potential, oxidative stress response, fate decisions, lineage commitment, redox state, cell signaling, and homeostatic capacity. Precise control of hypoxia in cell culture is vital for reproducible, pathophysiological cell data. However, there are important considerations when planning in vitro hypoxia experiments.


While hypoxic incubators have been available for decades, they are rarely used in cell culture experiments requiring oxygen control for several reasons. Pathophysiologic hypoxia, for many cells, is below 1% O2 and most O2/CO2 incubators are incapable of controlling extreme low oxygen levels. Beyond their narrow control range, multigas incubators consume excessive nitrogen gas and necessitate frequent cylinder exchanges. These recurrent N2 tank swaps pose logistical challenges but even more problematic is the adverse effect traditional O2 incubators have on generating robust hypoxic cell data. Since hypoxia-inducible factors respond quickly to changes in O2% , even brief exposures to room air oxygen (21% O2) which occur when opening/closing the incubator door and the subsequent recovery of O2 setpoint results in rapid degradation of HIF-1a and disruption of HIF protein biology. Multiple factors determine the appropriate O2% for each cell protocol and the limitation with using full-sized O2/CO2 incubators is their inability to simultaneously control more than one O2%. Since the oxygen level needed for each experiment can vary greatly , the single-function capability of full-sized hypoxic incubators makes them a poor option for carrying out multiple in vitro studies at the same time or even single studies that involve a small number of culture vessels.

P110 SC 1
Our most basic SubChamber System consists of a ProOx oxygen controller and a subchamber. The subchamber sits inside any existing temperature controlled incubator.


Protocols are different, each requiring a different piece to the hypoxia puzzle. Labs are different in their available equipment and size. Regardless of your specific situation, a clear and extensive upgrade path is ready to meet any hypoxia challenge you may encounter in your research.

Incubator Shuttle Chamber 3

Our SubChamber Systems make uninterruptible hypoxia easy. Inside a subchamber, you can move your culture undisturbed back and forth between incubator and any of our unique oxygen controlled glove boxes. Match O2 level in glove box with that of the culture chamber. Work with your cultures in the glove box without any disturbance in the O2 concentration.



Our SubChamber Systems are designed to fit right inside your existing incubator, adding valuable hypoxic functionality. Coupled with our versatile line of O2 controllers, SubChamber Systems provide many pieces: full range O2 control (0.1%-99.9%) for deep hypoxia, dynamic control of O2/CO2, new oxygen cousins NO and CO, and dissolved pericellular O2.

SubChamber Systems are sized to fit your cultures. Control hypoxia where needed, and save gas. Hypoxic conditions within the subchamber go undisturbed when the incubator door is opened. This provides hands-off uninterruptible conditions for your cells.

Subchambers can easily be transported to any of our glove boxes for aseptic processing.


The OxyCycler’s dynamic oxygen control provides the ability to do programmable physiologic modeling with ease.

EXPANSION PROTOCOL – Primary cultures and other low cell population density do best under low oxygen gas phase when they start, but when they grow, they are bound to need more. You can design any ramp up in oxygen, and consistently repeat it. With a click of the mouse.

ACUTE/INTERMITTENT HYPOXIA – Hypoxic stress can model components of many severe diseases such as heart attacks, strokes, asthma, or epilepsy. Frequency, duration and degree of drops are all adjustable.



Take the uncertainty and work out of flush box use. Our O2 controllers can provide accurate, consistent hypoxic conditions in your flush box. Rapid, measurable hypoxic conditions can be achieved and will lead to more consistent results for your research. Reduced handling requirements lower the risk of contamination. For uninterrupted cell processing manipulate your cells in any of our glove boxes.


Many hypoxia regulated genes are immediately turned off by brief exposures to ambient air. Handling your cultures in glove boxes with O2 control avoids this problem.

The I-Glove is a glove box for your incubator. Your incubator goes inside and the same conditions inside the incubator are now controlled outside the incubator. Any CO2 or O2/CO2 incubator can now provide deep hypoxia for your cells. Safely manipulate your cultures in the processing chamber and then return them to the incubator when finished. There is less danger of contaminating all of your cells as in standard glove boxes with shared incubation and processing space. Hypoxic conditions go undisturbed and virtually no gas is consumed. You can still add any of our SubChamber Systems to the incubator inside and benefit from the power and versatility they provide.


To eliminate the risk caused by incubating and processing cells in the same humid place, upgrade your workstation with an integrated incubation chamber module and continue to process in your workstation. Or integrate an additional processing module and continue to incubate in your workstation. Even add a microscope chamber with hypoxia control. All the other pieces can be integrated as well.


Where did all these upgrades come from? Over 30 years of meeting hypoxia challenges for thousands of hypoxia scientists worldwide.


After upgrading other equipment for 25 years, BioSpherix reinvented the concept of cell culture incubation and processing. The result is the XVIVO, the only closed-hood cell culture incubation and processing system which provides full-time, uninterruptible optimization of hypoxic conditions.

This closed-hood design takes the uncertainty created by random exposures to ambient air, its effect on gene expression and process control, out of the equation.


Available features include multiple, independent, fully-integrated incubators with dynamic programmable hypoxic control. Automatically run any hypoxic profile. Simultaneously run as few or as many protocols as you need. This feature accelerates your research. Dissolved pericellular O2 monitoring can be incorporated for better accuracy. Modular interconnected processing chambers can accommodate microscopes, centrifuges, cell sorters, bioreactors… whatever suits your needs. Your cells are never exposed to suboptimal conditions. Closed-hood incubation also greatly reduces the risk of contamination because the cells are never exposed to the outside environment or the technicians operating the equipment.

The Xvivo System Model X3 offers real-time monitoring with internet access to data logs and alarms. Full-time datalogging: a complete set of documents that prove the conditions your cells were exposed to. This may lead to a quicker and higher percentage of grant approvals.


You don’t have to compromise your research…or your budget. Get the capability you need today and the upgradeability for your research needs tomorrow. Whether it’s upgrading your current equipment, or creating your own BioSpherix modular hypoxic research system, we offer all the pieces to the hypoxic puzzle.


Other vendors have only one or two pieces of the hypoxia puzzle. Some have the hypoxia incubator. Some have the hypoxia glove box. Some have the hypoxia flush box. Some have a dissolved O2 analyzer. Only BioSpherix offers all the pieces. They’re integrated and modular. Start out small, and upgrade as needed. Every piece works perfectly with every other piece. Again, all the pieces to definitively solve the hypoxia puzzle.

Since 1982 we’ve been making unique tools for biomedical scientists. Our tools help you manipulate and control biologically active gases (O2, CO2, NO, CO, O3, H2S, etc.) in both in vitro and in vivo applications. We build controllers, chambers, partial systems, and complete systems. We can even “soup up” any existing third party equipment. If necessary, we can customize to exactly fit your needs. We guarantee solutions.

C Shuttle 4An I-Glove can enclose your existing O2/CO2 incubator, providing uninterrupted hypoxic cell incubation and processing.

Microscope 5

Aseptic hypoxia microscope chambers can be added to any hypoxia workstation.

Xvivo Drawers Steph 6

Cells inside the XVIVO are never exposed to the room. Any hypoxia challenge can be met.

Xvivo 7

BioSpherix XVIVO System A new concept in cell incubation and processing. Extreme modularity in all chambers and control functions allow you to configure for your needs now, then upgrade later if necessary. Unprecedented new abilities with no deadends.

Xvivo Gloves Incubators 8

Multiple integrated independent incubators open into controlled oxygen aseptic environment.