We provide a detailed explanation of the procedure of the histoculture drug response assay (HDRA) with 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) end point among several modified HDRA procedures. Fresh surgical specimens are cut into approx 1- to 2-mm3 pieces and put on a gelatin sponge infiltrated with culture medium containing a test drug. After incubation for 7 d, cell viability is assessed by the MTT assay. HDRA uses cancer tissue fragments with cells growing in three dimensions, with maintenance of intercellular contact and interactions with stromal cells. Therefore, it seems that HDRA can assess the sensitivity of tumor cells to anticancer drugs in conditions similar to those in vivo and, consequently, shows high prediction rate.