Cells grow in Complete Culture Media which contains all of t

Cells grow in Complete Culture Media which contains all of the nutrients required for cell growth, Complete culture media is made from basal media (in the case of RAW264.7 cells, it is called DMEM) to which Fetal Bovine Serum (FBS) and antibiotic are added, (i.e.. DMEM can be considered the \"diluents\" for Band antibiotic.) a. If you are making 200 ml of complete media, how much FBS (assume 100% solution) and 100X antibiotic would you add to achieve a final concentration of 10% FBS and IX antibiotic in DMEM? b. The concentration of glucose is 25 mM in most basal media (DMEM included). What is the final concentration of glucose in complete media? Are all of the other components (vitamins, amino acids, etc) of the basal media \"diluted\" to the same extent? Explain.

Solution

a) We will put our values in this formula. C₁V₁ = C₂V₂ where C₁ will be initial concentration and C₂ will be final concentration, V₁ will be initial volume and V₂ will be final volume. In order to make 10% FBS in 1000 ml basal media, we will place the values.

100 x a ml = 10 x 1000 ml , we will get value of a =100 ml, so we can say that 100 ml of FBS from 100 % solution will make 10% FBS in 1000 ml of basal media.

Similarly, 100X x a = 1000 ml x 1X , will get 10 ml, so we can say 10 ml of 100X antibiotic will make 1X of 1000 ml basal media.

Therefore 100 ml FBS+ 10 ml antibiotic +890 ml soultion (having, vitamins, amino acids, glucose etc) will make the basal media.

b) Glucose conc is 25 mM in 1000 ml basal media, so in 200 ml of complete media 5 mM will be the glucose concentration. (unitary method). Yes, akk the other components will be diluted similarly as DMEM (Dulbecco\'s Modified Eagle\'s Medium) contains higher concentration of these substancesd, which are further diluted in complete media of 200 ml.