Best Practices For Cell Culture Media Design And Processes

Sponsored by: Cell Culture Dish
Session ended: November 22nd, 2013
Expert: Paul J. Price, Ph.D., Media Design Consultant

Have you had the problem that a new cell line in your laboratory is not growing to your expectations or cells that had been growing well in your medium all of a sudden start looking sick and are going into apoptosis?

The classical cell culture medium consists of amino acids, vitamins and a source of energy, such as glucose, in a buffered salt solution. These formulations require further supplementation with a protein source such as serum. The classical media formulations were designed using cancer-derived cell lines and can be very sub-optimal for the growth of specialized cells, such as stem cell, recombinant cells and differentiated cells. In the absence of serum and serum proteins it becomes essential that you control for the osmolality, ammonia and the production of free radicals. As important as using the right medium is the proper handling of the both the media and the cells and together can be the difference between a successful or failed experiment.

This Ask the Expert Session is hosted by Paul J. Price, Ph.D., Media Design Consultant. Dr. Price has been a research scientist for over 50 years. Positions he has held include Branch Chief in the Center for Infectious Diseases at the CDC, and founder and Executive Vice-President of Hycor Biomedical.

If you have had a problem with an experiment or cells, take this opportunity to prevent either from crashing. Dr. Price has over 50 years of experience in cell culture and media design and has had the opportunity to make all of the mistakes and find ways to correct them.

Don’t miss the chance to ask your cell culture media related questions, session starts Today!


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Questions & Answers

We are having trouble with our VCC going down to half as we are moving adherent CHO cells to suspension. Any ideas on how media could help with the transition. We are using an off the shelf CHO media right now.

Going from adherent culture to suspension can be tricky, especially when going from a protein containing to a chemically defined (CD) formulation. The following procedure should work for you. Grow your cells in the medium that they have been adapted to and collect the supernatant(call it CM for conditioned medium) 2 to 3 days after […]» Read More

We are looking to streamline our process development and scale up. Do you think it is possible to use a single media for cloning, seed train expansion and production, with just minor modifications at each step? We are trying to eliminate using different media for each step of the process. Ideally we would just have one media, but if that is not possible then it would be nice to have one base media with adding a few components to each step as necessary

You probably could use one medium if you choose the correct one. For cloning you want to start with as small a volume as possible and the medium should have B12 as one of its vitamins. As cells grow they transfer nutrients and growth factors back and forth across their membrane. If the volume of […]» Read More

What kind of media do you recommend for transient transfection of HEK293 cells? Are there any key components to include or look for in media. Our cells aren’t doing well right now in our current media.

The medium will differ as will the transfection protocol dependent on if you are growing and transfecting adherent cells are cells in suspension. The life Technologies web-site has protocols on it for transfecting either type of cell. I just looked at a couple of them and they were pretty straight forward. I have never tried […]» Read More

If you are trying to remove or reduce serum in mesenchymal cells, what are some key components – supplements, growth factors, etc. that you need to include in your classic media.

To reduce serum in a classical medium you need to pick the right medium. GIBCO has OptiMEM 1 and several Advanced formulations such as Advanced DMEM:F12. You should be able to reduce the serum in these media to 2%. If you prefer to use a published claasical medium consider alpha MEM and supplement with ITS. […]» Read More

Yes we have. Cell is sensitive to the water, media lot and some contamination which is hard to identified. Sometimes we can’t find the reason which cause the bad growth and crashing.

Several different or combined problems may be causing your cultures to crash. A common cause of cells crashing is the destruction of the medium you are using by light. Both incadescent and fluorescent light can destroy the medium. The time it takes is dependent on the medium. A medium that is phenol-red-free and contains HEPES […]» Read More

A question regarding culture media sold as bulk pre-prepared liquid vs. powders for in-house preparation of media. Which is better performance-wise, liquid media from the manufacturer or liquids prepared from powders in-house? Everyone I’ve asked has noted that fresh liquid media from the manufacturer will inherently provide better cell culture performance vs. media prepared in-house from the ‘same’ comparably-fresh powder. Some report seeing >10% increase in yields using bulk liquid media vs. powder. Everyone presumes that the powder grinding process can only result in some loss of activity, if only from the heat involved; and that the culture media companies can simply make better, more consistent, finished liquid culture media than can beed in-house from powders, with the manufacturer simply knowing its products better, likely having heavier-duty and proprietary mixing technology/methods, etc.

The reason 1X liquid media from a manufacturer is often better then medium made in your lab from a DPM is the difference in the quality of water. The better commercial media companies monitor endotoxin and heavy metal contaminents in their water and have processes in place to control them. Water grows bacteria rapidly and […]» Read More

A question regarding culture media sold as bulk pre-prepared liquid vs. powders for in-house preparation of media. Which is better performance-wise, liquid media from the manufacturer or liquids prepared from powders in-house? Everyone I’ve asked has noted that fresh liquid media from the manufacturer will inherently provide better cell culture performance vs. media prepared in-house from the ‘same’ comparably-fresh powder. Some report seeing >10% increase in yields using bulk liquid media vs. powder. Everyone presumes that the powder grinding process can only result in some loss of activity, if only from the heat involved; and that the culture media companies can simply make better, more consistent, finished liquid culture media than can beed in-house from powders, with the manufacturer simply knowing its products better, likely having heavier-duty and proprietary mixing technology/methods, etc.

The reason 1X liquid media from a manufacturer is often better then medium made in your lab from a DPM is the difference in the quality of water. The better commercial media companies monitor endotoxin and heavy metal contaminents in their water and have processes in place to control them. Water grows bacteria rapidly and […]» Read More

We are trying to transition vero cells from serum containing to a commercially available serum-free media for vero, but are having trouble with the cells making the transition. Which steps would you recommend that we take to help the transition for the cells?

Cells, including VERO, work hard to make an environment for themselves allowing good viability and growth. When we discard the so-called spent or conditioned media, we are also discarding the growth and other factors the cells have produced for their survival. With primary neurons if you discard all of the spent medium they will go […]» Read More