The Research Of Guinea Pig Myopia Model And The TER Value Of Cultured Retinal Pigment Epithelial(RPE) Cells In Vitro

Object:Research the guinea pig myopia model induced by concave spherical lens, observe the transepithelial electrical resistance (TER) of cultured retinal pigment epithelial (RPE)cells in vitro,Explore and estable the stable expression time interval of TER value from the monolayer barrier model of RPE cell.Methods:Take30guinea pigs of the same batch of2-week-old tri-color guinea pigs assign into two groups randomly:the normal group (n=10) and monocular defocus model group (n=20), Referred to the defocus group, the focus group was established by wearing-10.00D Concave lens at front of the left eye of guinea pigs for two weeks, Measuring the refraction by dilating retinoscopy optometry and the axial length by A super-measuring instrument of the Two groups at2weeks and4weeks old eyes, then focus group guinea pigs were sacrificed, take the left eye RPE cells in primary culture, Take the cytokeratin and protein as markers, immunocytochemistry staining identified as positive.The cells were Transmitted to the second generation, inoculated on the surface of the microporous membrane, until the cell fusion as a single layer, culture in the containing1%serum concentration of culture-solution, Use resistance measuring instrument continuously detect and record the epithelial cell monolayer TER value.Result:1.the changes of refraction and axial length of defocus eye:The refraction of defocused eye myopia increased, focal group (-3.29±0.72D),normal group (+3.18±0.50D),The self-control group (+1.42±0.60D),showing significant differences between the difference (p<0.05); the axial length of defocused eye extended, the focus group (8.14±0.09mm) and the normal group (7.59±0.06mm), The self-control group (7.89±0.07mm)showing significant differences between the difference (p<0.05);2. RPE cells in vitro culture and identification from the defocused eye:The Defocus eye RPE cells cultured in vitro grow well, smooth-adherent, were expressed cytokeratin and S-100protein, the positive rate is100%, high purity of the cells. The Defocus eye RPE cells can preserve the inherent characteristics of the RPE cells,the passaged ability of RPE cells is good.3. the TER value detection from the defocus eye RPE cells:The TER value from defocus eyes of RPE cells increased with culture time extension, stabilize at about12days, to more than 57.18Ω/cm2, can maintain its stability for8days.Conclusion:1.The guinea pig myopia defocus model induced by the concave spherical can be successfully established;2.The growth and Passage capacity of myopia RPE in vitro of guinea pig from defocus eyes are good;3.The TER values of guinea pig myopia RPE in vitro from defocus eyes were stable with an epithelial cell monolayer structure, and the TER values can be maintained at least eight days, laid the basis of the experimental model for the next intervention research experiments.