Dulbecco s modified eagle medium

This study was approved by the Advanced Medical Research Center of Nara Medical University in Japan. Surgical procedures were conducted under routine sterile techniques. All procedures in thisstudy were performed by one neurosurgeon.
Gelatin was supplied by Nitta Gelatin Co. Ltd., Osaka. It was extracted from porcine skin to have an isoelectric point of 5. Phosphate-buffered saline (−), 25 wt% GA solution, 3-methyl-2-benzo-thiazoline hydrazone hydrochloride, and Dulbecco's modified Eagle medium were purchased from Wako Pure Chemical Inc., Osaka. Bovine serum albumin was purchased from Sigma-Aldrich (St. Louis, Missouri, USA). All reagents were used as obtained. Fibrin glue (Beriplast^®, CSL Behring, Victoria, Australia) was purchased from Wakenyaku Co. Ltd., Osaka. Doubly distilled water was used for all preparations. Gelatin and GA (1%) solutions were preheated to 45°C, and with the aid of an application device, these solutions were applied to the dura mater simultaneously with rubbing so that they mixed well, penetrated into the suture holes, and dried in 5 minutes.^{11) (link)}

Human melanoma cells (G361, SK-MEL5, A375s2, A375 and A2058) were purchased from the American Type Culture Collection (Rockville, MD, USA) and cultured at 37°C under conditions of 20% O₂ and 5% CO₂ in Dulbecco's modified Eagle medium (Wako, Osaka, Japan) containing 10% (v/v) fetal bovine serum, 20 mM 2-[4-(2-hydroxyethyl)-1-piperazinyl] ethanesulfonic acid, penicillin (50 units/ml) and streptomycin (50 μg/ml).

KFBS were obtained from chest and earlobe keloid tissues of two Japanese
patients. The protocol for tissue collection was approved by the ethics review
board at Kobe University Graduate School. Five or 6 pieces of the minced and
deepithelialized samples were placed on a 100-mm tissue culture dish (Iwaki,
Tokyo, Japan) and immersed in 2 mL Dulbecco's modified Eagle medium (DMEM; Wako
Pure Chemical Industries, Osaka, Japan) supplemented with 10% fetal bovine serum
(FBS; Nichirei, Tokyo, Japan), penicillin (50 U/mL), and streptomycin (50
µg/mL; MP Biomedicals, Illkirch, France). On the following day, 15 mL of
the medium was added to each dish. The culture medium was changed every 2 or 3
days until approximately 80% confluence was reached. The cells were passaged by
incubation at 37°C with 0.05% trypsin and 0.02% EDTA, and plated in culture
dishes.