Randy Yerden is the Founder and Chief Executive Officer of BioSpherix, Ltd. He created the Cytocentric Principles as a way to describe the fundamental biologic factors that drive the development of all of the products at BioSpherix. Here Chief Scientific Officer, Alicia Henn interviews Randy about how the Cytocentric Principles were formed. In part one of a multi-part series, we discuss applying cytocentric principles in future science research.
Keep the cells in log-phase growth for batch after batch. Rinse and repeat. The cells should be the same every time you need them. Seems like a simple thing, right?
But it never is that easy to get consistent cell growth. That’s why we spend so much time and money checking the cells. Do they still express the right markers? Before researchers even get to the biologically question at hand, a lot of space in scientific reports is dedicated to the simple question; “Are the cells being used the type of cell that they are supposed to be?”
We 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 . It can take up to 16 hours for cell culture to return to low oxygen levels in the incubator. 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.
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.
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.