The Application Of Collagen Gel Droplet Culture Drug-sensitivity Test In Pancreatic Cancer

Objective:To establish the CD-DST?collagen gel droplet embedded drug-sensitivity test?system.To explore the reliability of the system in drug-sensitivity test for pancreatic cancer cells,and then the application values of this system in the clinical treatment for the pancreatic cancer patients,which will assist in the selection of postoperative chemotherapy plan.Methods:1.Type I collagen solution,10X DMEM medium,and 1 nmol/L NaOH solution were mixed to establish a 3D cell culture system,of which the PH and osmotic pressure was suitable for the cell growth.2.The relationship between the number of cells in droplet and the OD value of the droplet picture was explored by detecting the OD values of the gel droplets which embedded different numbers of cells.3.The morphology and growth rate of human pancreatic cancer cell MiaPaca 2in the gel droplets were observed.4.The drug sensitivities of the MiaPaca 2 and MiaPaca 2/GZ cultured in 2D and 3D culture system were detected.5.The primary cells were separated from the surgical specimens of the 9 pancreatic cancer patients who underwent pancreatectomy from November 2017 to May 2018 in Pancreas Center of the Second Affiliated Hospital of Nanjing Medical University.The sensitivities of the primary cells to gemcitabine and fluorouracil were tested by CD-DST system.Results:1.A stable 3D culture system was established.The OD value of the gel picture was linearly related with the number of cells in the gel droplet.2.MiaPaca 2cells in this system are spherical with foot processes stretching peripherally.The cells grew in clusters after cultured for 6 days.3.The IC₅₀ concentration of gemcitabine to MiaPaca 2 and MiaPaca 2/GZ cells was 0.035?M?0.15?M in 2D system and 0.12?M?0.64?M in 3D system.The IC50 concentration of fluorouracil to MiaPaca 2cells was31.4?M in the 2D system and 89.4?M in 3D systems.Among the 9 specimens,there were 7 specimens from whom the primary cells were successfully separated and then tested for drug sensitivities in CD-DST system.The evaluate rate of the specimens was 77.8%.The drug sensitivities of primary cells from different patients varied.Conclusions:1.This study established a stable 3D culture system,in which the cells grow well.And it can be widely used in experimental studies at the molecular and cellular level.2.This system reliably detected the differences in drug sensitivities between common pancreatic cancer cells and drug-resistant cells.3.CD-DST system well reflected the differences of drug sensitivities to anti-tumor drugs in different pancreatic cancer patients.This system may play an auxiliary role in making the chemotherapy plan for pancreatic cancer patients,and the application value worth investigating in following clinical trials.