Optimized growth medium for primary culture of human oral keratinocytes.
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. 3H-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 micrograms/ml hydrocortisone, 5 micrograms/ml insulin, 10 ng/ml EGF, 2 micrograms/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 micrograms/ml), and hydrocortisone (5 micrograms/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.
Formanek
M
M
Department of Otorhinolaryngology, General Hospital, University of Vienna, Austria.
Millesi
W
W
Willheim
M
M
Scheiner
O
O
Kornfehl
J
J
D016475:3T3 Cells; D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D003470:Culture Media / Q000737:chemistry*; D006133:Growth Substances / Q000494:pharmacology; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D009061:Mouth Mucosa / Q000166:cytology
D003470:Culture Media; D006133:Growth Substances