Beriplast

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)}

Rat myoblasts L8 cells (American Type Culture Collection, Manassas, VA, USA; Cat. No. 95102434), which are commercially available and established from Wistar rats, were seeded onto temperature-responsive culture 6-well plates (UpCell 6 Multi-well plates; Cellseed, Tokyo, Japan; Cat. No. CS3004) at a density of 2 × 10⁵ cells/well. The cells were cultured in medium consisting of DMEM, high glucose (Thermo Fisher Scientific, Waltham, MA, USA; Cat. No. 11995–065), 10% fetal bovine serum (FBS; Thermo Fisher Scientific; Cat. No. 1606731) and 10 μg/mL Gentamicin (Fujiseiyaku, Tokyo, Japan) at 37 °C and 5% CO₂. The medium was replaced with fresh medium every 48–72 h. After 5 days of culture, confluent myoblasts on temperature-responsive dishes were transferred and left to sit at 20 °C for about 2 h, causing the myoblast sheets to detach spontaneously. The myoblast sheets were then washed 3 times with iced Hank's balanced salt solution containing calcium chloride and magnesium sulfate (HBSS+; Thermo Fisher Scientific; Cat. No. 14025–092) and fixed with fibrin tissue glue (Beriplast; CSL Behring, Malburg, Germany; Cat. No. 731141119).