Effect of subretinal injection of retinal pigment epithelial cells for retinal pigment epithelium in mice_Chinese Journal of Ocular Fundus Diseases

Authors：

ZhaoLiyu ,  KeXiaoyun

Department of Ophthalmology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China;

Corresponding author：

KeXiaoyun, Email: keyan96612@163.com

Keywords：

Retinal pigment epithelium/cytology; Cells, cultured; Animal experimentation

DOI：

10.3760/cma.j.issn.1005-1015.2016.01.014

Video：

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Objective To observe the effect of subretinal injection of retinal pigment epithelium (RPE) cells for RPE in mice. Methods A total of 30 postnatal day 7 C57BL/6J mice were randomly divided into normal mice group, OIR model group and OIR model cell transplanted group, 10 mice in each group. The OIR model was induced in mice of OIR model group and OIR model cell transplanted group. The RPE cells were subretinal injected into the RPE of mice in OIR model cell transplanted group. At 20 days after the injection, the RPE thickness was evaluated by fluorescence microscope. The expression of RPE65, Bestrophin and zonula occludens-1 (ZO-1) were estimated by Western blot and real-time quantitative PCR (RT-PCR). Results The thickness of RPE in OIR model mice was thinner than that in normal mice; the thickness of RPE in OIR model cell transplantation mice was significantly thicker than that in the OIR model mice. The results of Western blot and RT-PCR indicated that the differences of protein (F=8.597, 18.864, 25.691) and mRNA expression (F=39.458, 11.461, 34.796) of RPE65, Bestrophin, ZO-1 were statistically significant between OIR model group and OIR model cell transplanted group (P < 0.05). Conclusions Subretinal injection of RPE cells can promote RPE thickening. RPE65 and Bestrophin protein relative expression levels increased, ZO-1 protein relative expression levels reduced; mRNA expression levels of RPE65, Bestrophin and ZO-1 genes increased.

Citation： ZhaoLiyu, KeXiaoyun. Effect of subretinal injection of retinal pigment epithelial cells for retinal pigment epithelium in mice. Chinese Journal of Ocular Fundus Diseases, 2016, 32(1): 58-61. doi: 10.3760/cma.j.issn.1005-1015.2016.01.014 Copy

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1. 胡诞宁.胚胎干细胞衍生的视网膜色素上皮细胞移植治疗眼病.中华细胞与干细胞杂志:电子版, 2014, 4(2):104-108. DOI:10.3877/cma.j.issn.2095-1221.2014.02.005.Hu DN. Therapy for retinal degenerative diseases by human embryonic stem cells derived retinal pigment epithelium transplantation[J]. Chin J Cell Stem Cell (Electronic Edition), 2014, 4(2):104-108. DOI:10.3877/cma.j.issn.2095-1221.2014.02.005.
2. Wert KJ, Skeie JM, Davis RJ, et al. Subretinal injection of gene therapy vectors and stem cells in the perinatal mouse eye[J]. J Vis Exp, 2012, 25(69): 4286. DOI:10.3791/4286.
3. Paris L, Tonutti L, Vannini C, et al. Structural organization of the tight junctions[J]. Biochim Biophys Acta, 2008, 1778(3):646-659.
4. Wright CB, Redmond TM, Nickerson JM.A history of the classical visual cycle[J].Prog Mol Biol Transl Sci, 2015, 134:433-448.DOI:10.1016/bs.pmbts.2015.06.009.
5. Müller C, Más Gómez N, Ruth P, et al. CaV1.3 L-type channels, maxiK Ca²⁺-dependent K⁺ channels and Bestrophin-1 regulate rhythmic photoreceptor outer segment phagocytosis by retinal pigment epithelial cells[J]. Cell Signal, 2014, 26(5):968-978. DOI:10.1016/j.cellsig.
6. Rizzolo LJ, Peng S, Luo Y, et al. Integration of tight junctions and claudins with the barrier functions of the retinal pigment epithelium[J]. Prog Retin Eye Res, 2011, 30(5):296-323. DOI: 10.1016/j.preteyeres.2011.06.002.
7. Smith LE, Wesolowski E, McLellan A, et al. Oxygen-induced retinopathy in the mouse[J]. Invest Ophthalmol Vis Sci, 1994, 35(1):101-111.
8. Zhang L, Tang S, Huang B, et al. The retinoid acid injected into sub retinal space couldn't reverse the retinal degeneration of mouse[J]. Eye Science, 2005, 21(2):116-118, 131. DOI:10.3969/j.issn.1000-4432.2005.02.014.
9. Fischer S, Wobben M, Marti HH, et al. Hypoxia-induced hyperpermeability in brainmicrovessel endothelial cells involves VEGF-mediated changes in the expression of zonula occludens-1[J]. Microvasc Res, 2002, 63(1):70-80.
10. Fischer S, Wiesnet M, Marti HH, et al. Simultaneous activation of several second messengers in hypoxia-induced hyperpermeability of brain derived endothelial cells[J].J Cell Physiol, 2004, 198(3):359-369.

Chinese Journal of Ocular Fundus Diseases

Effect of subretinal injection of retinal pigment epithelial cells for retinal pigment epithelium in mice

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