Table 3

Media composition in the different steps of cell culture optimization

Initial media testing

Basic media

Supplementation


DMEM +10% FCS

-

DMEM/Ham's F-12 (1:1) +10% FCS

-

Ham's F-12 +10% FCS

-

McCoys-5A +10% FCS

-

Medium 199 +10% FCS

-

Media conditioning and additives

Basic media

Supplementation

Ham's F-12 +10% FCS

-

Ham's F-12 +10% FCS, cond.

-

Ham's F-12 +10% FCS, cond.

+ EGF

Ham's F-12 +10% FCS, cond.

+ insulin

Ham's F-12 +10% FCS, cond.

+ hydrocortisone

Ham's F-12 +10% FCS, cond.

+ EGF, insulin

Ham's F-12 +10% FCS, cond.

+ insulin, hydrocortisone

Ham's F-12 +10% FCS, cond.

+ EGF, hydrocortisone

Ham's F-12 +10% FCS, cond.

+ EGF, insulin, hydrocortisone

Differentiated cell culture

Lower compartement

Upper compartment

Ham's F-12 +10%FCS, cond., EGF, insulin

a) Ham's F-12 +10%FCS, cond., EGF, insulin

b) no medium (air-liquid interface)


DMEM Dulbecco's MEM medium, FCS fetal calf serum, cond. conditioned. Concentration of the supplements: 10 ng/mL murine EGF, 1 μg/mL porcine insulin, 0.5 μg/mL hydrocortisone. All growth media were supplemented with 100 U/mL penicillin/100 μg/mL streptomycin, 50 μg/mL gentamycin, 1 μg/mL amphotericin B, 10 μg/mL reduced glutathione and 10 μg/mL ascorbic acid.

Miessen et al. BMC Veterinary Research 2012 8:31   doi:10.1186/1746-6148-8-31

Open Data