Barrier function of porcine retinal pigment epithelial cells cultured in vitro / 中华眼底病杂志

ABSTRACT

Objective To investigate the blood-retinal barrier(BRB)function of porcine retinal pigment epithelial(RPE)cells cultured in vitro. Methods Primary porcine RPE cells were cultured,and the third generation were inoculated in a microporous filter with the filter membrane of polyvinylpyrrolidone(PVP)-free polycarbonate membrane.After 1,2,3 and 4 weeks of culture,the surface of filter membrane was observed by light microscope,and after 2 weeks of culture,the section of filter membrane was observed by light microscope and transmission electron microscope.Transepithelial electrical resistance(TER)was detected and the permeability was measured with fluorescein sodium(FS)and horseradish peroxidase(HRP). Results Primary porcine RPE cells were cultured successfully.RPE cells converged1week after inoculation; 2 and 3 weeks after inoculation,the density of RPE cells did not changed obviously; 4 weeks after inoculation,the density of RPE cells decreased.The characteristics of polarized growth of monolayer were found in RPE cells on the surface of filter membrane; 2 weeks after inoculation,the TER of RPE cells was(97.44?11.36)?/cm~2 which maintained till the 3rd week after inocubation.After incubated for 30 minutes,only 0.27% of FS and 0.17% HRP reached the inferior filter membrane,and the permeability rate of SF with low molecular weight was higher than which of HRP with high molecular weight. Conclusions The filter with PVP-free polycarbonate membrane may be used to set up the model of RPE cells with polarized growth of monolayer and investigate the barrier function of RPE cells.