Chemically Defined Eagle’s Minimal Essential Medium (CD-EMEM)

Description

Background

PurMa™ Chemically Defined Eagle’s Minimal Essential Medium (CD-EMEM) is a proprietary formulation used to grow cells in vitro without any undefined or animal-derived components. Moreover, CD-EMEM contains known quantities of pure chemicals, including amino acids, vitamins, salts, sugars, and more importantly a proprietary combination of synthetic growth factors.

It is absolutely crucial to mention that the PurMa™ Growth Supplement for each conventional media such as DMEM, IMDM, RPMI-1640… (Base Media ) are slightly different in combination, amount of several growth factors, ions, chemicals,  pH with particular adjustments needs to be done, based on the environments which base media provides.

This provides a consistent and controlled environment for cell growth and research. Noticeably, CD-EMEM is essential for applications that require reproducibility and regulatory compliance as well as for studying specific cellular processes, without extraneous variables. Furthermore, CD-EMEM is designed for custom formulations tailored for various cell types.

Important Notes:

It is crucially important to have a gradual transition from media containing FBS to chemically defined cell culture media. Follow these steps:

• Starting Point: 10% FBS+ (control) (you need two controls, one to be left in only 10% FBS and continue monitoring for comparing with 0% FBS + PurMa^TM Chemically Defined Medium. The second one is chemically defined medium + 10% FBS, which is going through the reduction of FBS process)
• 48 hours later: 8% FBS in + PurMa^TM Chemically Defined Medium
• 48 hours later: 6% FBS in + PurMa^TM Chemically Defined Medium
• 48 hours later: 4% FBS + PurMa^TM Chemically Defined Medium
• 48 hours later: 2% FBS + PurMa^TM Chemically Defined Medium
• 48 hours later: 1% FBS + PurMa^TM Chemically Defined Medium
• 24 hours later: 0% FBS and only PurMa^TM Chemically Defined Medium

Importantly, for each change, spin down and fully discard the media and replace it with the next lower percentage of FBS.

Noticeably, you don’t have to start with a huge number of cells (~500,000 cells/ml). Additionally, we strongly recommend you start with more than one plate as some of the plates might get contaminated.

So, once the cells are kept in chemically defined Media, you should always keep them in the same chemically defined medium from the same vendor and the same formulation, even for freezing (see below).

Usually, after you get your cells adapted to PurMa^TM Chemically Defined Medium you need to make several plates and freeze down several vials. The mentioned cells are adapted to chemically defined medium and when you thaw them in the first week, in addition to PurMa^TM Chemically Defined Medium, you need to add 10% PurMa^TM CD Booster (Cat# P3C10144).

Formulation

The CD-EMEM includes a base formulation including L-Alanine -L Glutamine : Base formulation of Chemically Defined EMEM , P3C310110

Additionally, CD-EMEM contains a proprietary synthetic cocktail of specific growth factors, hormones, and other nutrients which makes it independent of any serum and any trace of animal products.

References

1. Up-regulation and increased phosphorylation of protein kinase C (PKC) delta, mu and theta in the degenerating rd1 mouse retina.   Azadi S,et al,Mol Cell Neurosci. 2006 Apr;31(4):759-73. doi: 10.1016/j.mcn.2006.01.001. Epub 2006 Feb 28.
2. Immunization and culture of rainbow trout organ sections in vitro. Anderson DP, Dixon OW, Lizzio EF. Vet Immunol Immunopathol. 1986 Jun;12(1-4):203-11. doi: 10.1016/0165-2427(86)90124-8.
3. Human plasma and amino acids as moderators of uptake and metabolic consequences of antifolates in WIL-2 and human leukemia cells. Fyfe MJ, Sedwick WD, Brown OE, Laszlo J. J Natl Cancer Inst. 1981 Mar;66(3):445-51.

Learn more: What Is EMEM?