The Establishment Of Collagen Gel Droplet Embedded-culture Drug-sensitivity Test With Rat Tail Collagen And Its Application To Uterine Cervix Cancer

Objective:To establish the collagen gel droplet embedded-culture drug-sensitivity test (CD-DST) with rat tail collagen and discuss the applied value of the technique above, and then to provide a available route for the scheme of clinical chemotherapy.Methods:The first, to construct three-diamensional formwork of cell culture in vitro, Id est, Hela cell suspension was added to a collagen solution which was a mixture of self-made rat-tail collagen, ten fold concentrated F-12 medium and 0.5 mol/L NaOH solution, and made collagen-cell mixture coagulate. Hela cells were cultured in the formwork, compared morphology with two-dimensional cultivation; then, to set up collagen gel droplet drug sensitivity test, this method consists of collagen gel droplet embedded culture step, exposure anticancer drugs, a drug-free culture step, and evaluation of anticancer effect by image analysis. drug sensitivity is detected by MTT and CD-DST, and evaluate the feasibility of this method; the final, applying this method to chemosensitivities test of twelve biopsy specimens in vitro.Results:1. Cell-collagen mixture could coagulate and form rat-tail collagen gel drop at 37℃in a CO2 incubator for 15 minutes. Hela cells cultured in three-diamensional formwork grew better, clonally proliferated, cells dispersed in it, appearance of cells cultured two-dimensionally was polygon,dispersal and individual.2. We have established the collagen gel droplet embedded-culture drug-sensitivity test (CD-DST) with rat tail-collagen. Comparing CD-DST with MTT, inhibition ratio with CD-DST was higher than that with MTT on identical drug concentration, but two results of the drug sensitivity had better consistency.3.12 cases of uterine cervix cancer had been detected by the CD-DST,6 cases could be separated tumor cells and be used for drug sensitive test, sample appreciation rate was 50% (6/12); drug sensitive of the 6 cases to 4 chemotherapeutics were different.Conclusions:1. This study establishes three-diamensional formwork of cell culture in vitro, It provides a visual and reliable in vitro model of observing the cells growth status and proliferation in the simulation of human environment similar, and lay a foundation in the future for the growth of cancer research in vivo, transfer, resistance and anticancer drug sensitivity test,etc.2. The collagen gel droplet embedded-culture drug-sensitivity test (CD-DST) with rat-tail collagen could be used for screening of drug-sensitivity test in vitro and reflect the heterogeneity of individuals to drug-sensitivity of tumors. Further more, the amount of cells for detecting is few (3×10 cells/hole) and enlarging the extension of clinical application. It is feasible for drug-sensitivity test in vitro.