Inner blood-retinal barrier in vitro model of Müller glial cells to influence tight junction.docVIP

 PAGE \* MERGEFORMAT 15 Inner blood-retinal barrier in vitro model of Müller glial cells to influence tight junction 【Abstract】 In vitro inner blood-retinal barrier (iBRB) models and to study retinal Müller glial cells (RMGC) of the tight junction (TJ) effects. Methods: Immunomagnetic method of purification and primary culture of rat retinal microvascular endothelial cells (RMEC), using modified enzyme digestion method of cultivation RMGC, RMEC and RMGC on both sides of the porous membrane co-culture model of the establishment of iBRB by phase-contrast microscope and Transmission electron microscopy (TEM) observation of two kinds of cell growth and TJ forms. Endothelial cells cross-resistance (TER) measurements and TJ-associated protein ZO-1 staining and protein electrophoresis to evaluate changes in TJ. Results: RMEC and RMGC can grow normally in the membrane, RMGC the close connection between the. TEM observations of high TER values of the samples adjacent RMEC forming a typical TJ structures. RMEC layer TER reached at 8d highest (86.3 ± 7.9) Ω * cm2, then remained relatively stable. RMGC exist in co-culture when TER 7d group (97.6 ± 3.6) Ω * cm2, is about 7d a two-fold when the RMEC group, 8 d peak (113.5 ± 5.3) Ω * cm2. The iBRB model development process, ZO-1 expression intensity increased with time. Conclusion: RMEC and RMGC of the co-culture can be established in vitro iBRB model, RMGC speed RMEC maturity between the TJ. 【Key Words】 Retinal 0 Introduction The retina in order to effectively perform its functions, needs a stable internal environment, the internal environment to maintain the stability depends on the inner blood-retinal barrier. Inner blood-retinal barrier by retinal microvascular endothelial cells (RMEC), RMEC the close connection between (TJ), basement membrane, pericytes, glial cell foot processes and very narrow extracellular space composed of a complex, is exist in the retinal nerve layer of blood vessels