Culturing rats lung fibroblasts and establishment of acute inflammation model in vitro

Objective: To study the method of culturing rats lung fibroblasts and establishment of acute inflammation model in vitro.Methods: After the lung fibroblasts was isolated and purified with trypsin and different adherence,the cells began to be cultured.The nature of the cultures was identified by immunofluorescence.The acute inflammation model was established with LPS.Cells were divided into control group,0.1μg/mL group,1μg/mL group and 10μg/mL group.The value of OD is determined after the cells exposed to LPS for 3 h,6 h and 9 h with MTT.Results: After the fibroblasts were cultured for 24 h,long spindle cells climbed out surrounding the tissue were seen under microscope.It growed rapidly at 72 h and got fusional in 4 days.Through lung fibroblast cells were measured by immunofluorescence,the vimentin was positive and CD31 was negative.In the groups of different LPS concentration,OD values of 1μg/mL LPS group were higher than that in other groups(P 0.05).In 1μg/mL LPS group,the OD values measured at 6 h were higher than that of other time periods(P 0.05),the difference was statistically significant.Conclusion: The experimental method can culture the rats lung fibroblasts in vitro successfully.It is suitable to establish the acute inflammation model with the concentration of LPS 1μg/mL for 6 h.