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!
Questions & Answers
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 MoreYou 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 MoreThe 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 MoreFor best results you probably want to grow your cells in suspension and in a CD-medium. The various CD media are cell type specific as different cells utilize the amino acids at different rates. You also have to decide if you are going with fed-batch or perfusion. You would be better off buying a specialty […]» Read MoreYou need to be aware that different nutrients are used at different rates and what is overly utilized is cell specific. With CHO cells glutamic acid, aspartic acid and cystene are self limiting and need to be part of a perfusion medium. Glutamine or glutamine dipeptide needs to be added at about 0.5 mM in […]» Read MoreTo 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 MoreSeveral 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 MoreThe 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 MoreThe 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 MoreIt not the cost of the ingredients that dictate the product cost but the labor involved. A few of the ingredients in a CD formulation are costly (i.e. monothioglycerol) but the concentrations in the 1X formula are so low as to not be a significant factor. A CD medium is a very complicated medium because […]» Read MoreCells, 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