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Optimized growth medium for primary culture of human oral keratinocytes

Formanek, Michael ; Millesi, Werner ; Willheim, Martin ; Scheiner, Otto ; Kornfehl, Johnannes ;
  • 초록  

    Abstract The effect of different media additives in defining optimal growth conditions for primary cultures of human oral keratinocytes was studied. A cocultivation technique with irradiated Swiss-3T3-fibroblasts in 96-well plates enables the comparison of additives for primary keratinocyte cultures derived from one patient. 3 H-labeled thymidine uptake showed no growth or growth inhibition with adenine, choleratoxin or transferrin compared to basal medium (Dulbecco's modified Eagle's medium (DMEM) and 10% fetal calf serum). Among single additives, 5 μg/ml hydrocortisone, 5 μg/ml insulin, 10 ng/ml EGF, 2 μg/ml bovine pituitary extract, and 10 −9 M triiodothyronine showed the greatest capacity to promote keratinocyte growth. With all possible combinations of additives, maximum stimulation was found with a combination of EGF (10 ng/ml), insulin (5 μg/ml), and hydrocortisone (5 μg/ml); none of the other combinations were more effective. Our data indicate that in short-term cultures (up to 5 days) various media additives described in the literature are not necessarily required in this system of primary culture of human oral keratinocytes.


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  • 주제어

    mucosa .   media additives .   proliferation assay .   growth supplements.  

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