Nutrient mixture f 12 ham ham s f12

For the experimental animal diets, the vitamin mixture, mineral mixture, cellulose powder, and corn starch were purchased from the Oriental Yeast Co. (Tokyo, Japan). At the same time, the soybean oil was acquired from the Nacalai Tesque (Tokyo, Japan), and the corn oil and sucrose were acquired from the Fujifilm Wako Pure Chemical Corporation (Osaka, Japan). Recombinant human IGF-I was obtained from Astellas Pharma Inc. (Tokyo, Japan). Nutrient Mixture F-12 Ham (Ham’s F12, #F6636, Sigma, St. Louis, MO, USA), William’s E medium (#W4125), 10× Earle’s buffered salt solution (EBSS), 100× MEM vitamin solution, and fetal bovine serum (FBS) were purchased from the Sigma Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM) was purchased from the Nissui Pharmaceutical Co. (Tokyo, Japan). Human recombinant growth hormone (Growject^®) was obtained from the JCR Pharmaceuticals (Hyogo, Japan). Other reagent-grade chemicals used in this study were commercially available.

The human cervical carcinoma cells (HeLa) and laryngeal carcinoma cells (Hep-2) were grown in a 1:1 mixture of DMEM and Nutrient mixture F-12 Ham (Ham’s F-12, Sigma) supplemented with 10 % FBS (Life Technologies, Karlsruhe, Germany), penicillin (100 μg/ml, Sigma), and streptomycin (100 μg/ml, Sigma). The cultures were kept at 37 °C in humidified atmosphere of 95 % air and 5 % CO₂.