Chemically Defined Basal Medium Eagle (CD-BME) in Earle’s Buffer

Description

Background

PurMa™ Chemically Defined BME in Earls’ buffer (CD-MEM- EBSS) is a proprietary formula used to grow cells in vitro without any undefined or animal-derived components. Moreover, CD-MEM- EBSS Medium 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 the base media provides.

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

Important Notes

It is crucially vital for 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^™ Chemically Defined Medium. The second one is chemically defined medium + 10% FBS, which is going through the reduction of the FBS process.)
• 48 hours later: 8% FBS in + PurMa^™ Chemically Defined Medium
• 48 hours later: 6% FBS in + PurMa^™ Chemically Defined Medium
• 48 hours later: 4% FBS+ PurMa^™ Chemically Defined Medium
• 48 hours later: 2% FBS+ PurMa^™ Chemically Defined Medium
• 48 hours later: 1% FBS + PurMa^™ Chemically Defined Medium
• 24 hours later: 0% FBS and only PurMa^™ Chemically Defined Medium

Importantly, for each change, spin down and fully discard the media and replace it with a 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 medium, 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^™ 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™ Chemically Defined Medium, you need to add 10% PurMa^™ CD Booster (Cat# P3C10144).

Formulation

The CD-MEM- EBSS Medium includes a base formulation including L-Alanine -L Glutamine: Base formulation for CD BME in EBSS P3C310891

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

References

1. Improved chemically defined basal medium (CMRL-1969) for primary monkey kidney and human diploid cells. Healy GM, et al. Appl Microbiol. 1971 Jan;21(1):1-5. doi: 10.1128/am.21.1.1-5.1971.
2. Nutritional requirements for the production of herpes simplex virus. I. Influence of glucose and glutamine of herpes simplex virus production by HeLa cells. LEWIS VJ Jr, SCOTT LV. J Bacteriol. 1962 Mar;83(3):475-82. doi: 10.1128/jb.83.3.475-482.1962. PMID: 14464909