Cytocentric-Blog

What Can I Do To Get the Best Results from the Non-Physiologic Cell Handling Conditions of a Room Air BSC?

bp28 images01We posted previously about how the Biology of HIF Proteins Impacts the Outcome of Your Experiments. Of course, full-time control of cell handling conditions including oxygen, CO2, and temperature is the in vivocondition and this is best in vitro as well. Changes in HIF protein levels have been found within minutes of oxygen changes [1]. It can take up to 16 hours for cell culture to return to low oxygen levels in the incubator[2]. Cells Need Optimization and the longer the cells are out of optimal conditions, the more likely they are to be affected by those conditions. Adapting to Cytocentric cell culture techniques means that routine passaging of your cells is not business as usual.

Oxygen Terminology

"Should We Call Physiologic Oxygen Hypoxia, Normoxia, Physioxia, or Something Else?"

bp029 image001

Being able to replicate research results is dependent on having the same conditions each time. Continue reading to learn about the factors to consider when classifying room air oxygen levels.

How we describe oxygen in the microenvironment is important.

The use of physiologically relevant oxygen for in vitro cell culture is increasingly essential as cells grown in vitro become more clinically important. Oxygen levels are a critical cell parameter, just like carbon dioxide or temperature. Hyperoxia simply means too much oxygen, Normoxia means normal amounts, and Hypoxia means too little.

What Oxygen Level Does My Cell Culture Actually Experience?

image002

Read about 3 protocol factors that affect your cell cultures below.

Well, it depends upon your cell culture protocol.

Just like in the perfect cookie recipe1, there are many factors in cell culture that affect your results. There is one set of ingredients or reagents that we don’t often think about when placing cultures into the incubator and shutting the door: the gasses. Even the 5% CO2 reading on the incubator isn’t what your cells are actually experiencing; not until the cell cultures have equilibrated with the incubator.

That takes time: time that your cells are out of optimum conditions.

The Biology of HIF Proteins Impacts the Outcome of Your Experiments in Physiologic Oxygen:

Considerations for protocol design

We recently posted an article “What Oxygen Level Should I Use for My Cells In Vitro?” in response to questions that we have fielded from customers getting started in cytocentric research. To expand on that topic, we are posting here a few more resources for information on the mechanism of how changes in oxygen levels impact cells in culture.

Mesenchymal Stem Cells for Cellular Therapies and Regenerative Medicine: 

Cytocentric oxygen control makes sense from the molecular level through to the bottom line.

Mesenchymal stem cells (MSC) have shown tremendous promise as a cellular therapy for indications as varied as arthritis [1], diabetes [2], cardiac disease [3], wound repair [4], graft-versus-host disease [5], ALS [6], spinal cord injury [7], even infectious disease [89]. MSC also have the potential to transdifferentiate into multiple cell lineages for regenerative medicine applications [10]. But for MSC to live up to that promise, the best conditions for expanding and studying these critical cells in the laboratory must be established.