|Budget Amount *help
¥2,000,000 (Direct Cost : ¥2,000,000)
Fiscal Year 1995 : ¥700,000 (Direct Cost : ¥700,000)
Fiscal Year 1994 : ¥1,300,000 (Direct Cost : ¥1,300,000)
An assay method was newly developped for evaluation of LPS-resistance : Both 10 mug/ml cycloheximide and 10 ng/ml LPS induced the severe cell damages in an LPS-sensitive macrophage-like cell, JA-4, a subline of J774.1, andafter incubation at 37ﾟC for 4th.However, LPS1916 cell line, a highly resistant cell line to LPS,was not damaged at all, even after incubation with cycloheximide and 1 mug/ml LPS.This method was sensitive and rapid for estimation of LPS-resistance, as compared with a cell growth assay or a colony-formation assay.
Trials to transfect DNA to JA-4 cells were not successful with calcium phosphate method, electro-poration method or lipofection method.A preliminary experiment is being performed by using another macrophage-like cell line, RAW 264.7, which is sensitive to LPS,as a receipient of the cDNAa from LPS1916 as well as JA-4 cells, because a recent report from other laboratory suggested that RAW 264.7 cells expressed luciferase gene after transfection of the gene.