Cytocentric Blog

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Cytocentric Visionaries: Ian Mudway, King's College, UK

Part Three: We Will Know We Have Gotten Through When People Complain About the Difficulties of Publishing Data Generated under Atmospheric Oxygen

 

In Part Two, Dr. Henn talked with Dr. Ian Mudway about difficulties in getting publishers and the greater scientific community to embrace his important findings. Here, they discuss overcoming scientific inertia and paradigm shift.

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Cytocentric Visionaries: Ian Mudway, King's College, UK

Part Two: Taking a Career Risk for Better Cell Culture and Better Science

 

In Part One, Dr. Henn talked with Dr. Ian Mudway about his recent publication , “Quantifying the magnitude of the oxygen artefact inherent in culturing airway cells under atmospheric oxygen versus physiological levels.”

Here, they discuss barriers to change in the scientific community.

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Cytocentric Visionaries: Ian Mudway, King's College, UK

Part One: What Are the Fundamental Things That We Absolutely Have To Get Right?

Alicia Henn, Chief Scientific Officer, BioSpherix

Ian Mudway is a Lecturer in Respiratory Toxicology at King’s College in London. He has been active in air pollution research, participating in Air Quality conferences, speaking out on BBC World News and in the Guardian newspaper, and advising the World Health Organization.

Here, Dr. Henn talks with Dr. Mudway about his recent publication , Quantifying the magnitude of the oxygen artefact inherent in culturing airway cells under atmospheric oxygen versus physiological levels.”

 

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Mini-Review: Cord Blood HSC and Physiologic Oxygen

 

At what oxygen level should I culture my umbilical cord-derived hematopoietic stem cells? It’s not a question you will hear on Jeopardy, but it is a critically important question.

Hematopoietic stem cells from umbilical cord blood (CB-HSC) have been shown to be beneficial for treating leukemia1,2, neuroblastoma3, and is of great interest for other disease applications. NIH-funded clinical trials listed on clinicaltrials.gov include studies of CB-HSC for immunodeficiencies, autoimmune diseases, sickle cell, cerebral palsy, complications of prematurity, and stroke. However, individual cord blood units often do not have the cell yield needed for a full transplant, requiring the pairing of HLA- partially matched cord blood units for a single transplant4. This presents obvious drawbacks in limiting the numbers of patients that can be treated.

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What is the O2 Level in the My Cell Culture Incubator? (Hint: It’s Not 21%)

Because room air is assumed to be near 21% oxygen, researchers often report that their room air incubator is as well. However, if you would think about it for a minute, you would realize that this is impossible.

 We charge a traditional open room cell culture incubator with CO2 , raising it to 5% or more, to maintain the pH of carbonate buffer-based cell culture media. Room air contains only 0.03 - 0.06% CO2 (or sometimes a little more if your ventilation is poor). Every time the inner incubator door is opened and closed, CO2 is infused to replace what is lost to the room. This necessarily drives the percentage of oxygen down inside the incubator.

High humidity also drives incubator oxygen levels down as water vapor can take up about 8% of the gas volume in an incubator with an open water pan. We have had customers with high oxygen applications report to us that they could not drive oxygen levels higher than 87% with 5% CO2 and an open water source.

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