Protective effect of retinal stem cell transplantation on retinal ganglion cells in glaucoma

doi:10.3969/j.issn.2095-4344.2016.28.015

Abstract

Abstract:

BACKGROUND: Stem cell transplantation is a new method for blinding eye disease. But there is a lack of research about the protective effect of retinal stem cell transplantation on retinal ganglion cells in glaucoma.
OBJECTIVE: To explore the protective effect of retinal stem cell transplantation on retinal ganglion cells of rats with glaucoma.
METHODS: Forty-five Sprague-Dawley rats were randomly divided into three groups (n=15 per group) including control, model and retinal stem cell transplantation groups. Rat models of glaucoma were prepared in the latter two groups, and at 7 days after modeling, rats in the three groups were given intravitreal injection of 1 mL retinal stem cells (5x10⁶ cells), the same amount of PBS, and no treatment, respectively. Subsequently, relative indicators were detected at 2 weeks after transplantation.
RESULTS AND CONCLUSION: The expressions of brain-derived neurotrophic factor and insulin-like growth factor I protein as well as the number of retinal ganglion cells were the highest in the control group, followed by the retinal stem cell transplantation group model group, and the lowest in the model group (P < 0.05). The number of apoptotic retinal ganglion cells in model group was significantly higher than that of control group (P < 0.05), and which in the retinal stem cell transplantation group was significantly lower than that in the model group (P < 0.05), but higher than that in the control group (P < 0.05). These results suggest that retinal stem cell transplantation for rat glaucoma can exert a protective effect on retinal ganglion cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Retina, Stem Cell Transplantation, Glaucoma, Retinal Ganglion Cells, Tissue Engineering

CLC Number:

R394.2

Gu Zhi-min, Zhou Li-xiao, Qi Ruo . Protective effect of retinal stem cell transplantation on retinal ganglion cells in glaucoma[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(28): 4203-4209.

Figures/Tables 1

2.1 视网膜干细胞的形态原代培养第3天出现细胞球，形状为类圆形，原代培养第5天，细胞球增多增大，折光性增强，形成立体感比较强的神经球，培养第7天以后神经球增大不明显。视网膜干细胞培养至第7天时开始传代培养，传代培养第3天出现细胞克隆，呈小圆形，第2代细胞培养24 h出现细胞团(图1)。 2.2 视网膜干细胞的鉴定免疫荧光染色鉴定结果显示：培养细胞中96%以上细胞为Nestin阳性表达细胞(图2)。 2.3 大鼠青光眼模型的鉴定在术后第1天有20只大鼠实验眼出现眼结膜轻度水肿，11只出现角膜轻度水肿，6只出现前房房水混浊，术后第7天时所有症状均消失，实验眼均未出现晶状体浑浊。30只对照眼均未出现异常。由表1可以看出，30只实验眼在术后第7天和第21天时的眼内压均明显高于对照眼(P < 0.05)，表明青光眼大鼠模型均建造成功。 2.4 各组大鼠视网膜中脑源性神经营养因子和胰岛素样生长因子Ⅰ蛋白的表达由表2可以看出，模型组脑源性神经营养因子和胰岛素样生长因子Ⅰ蛋白的表达均明显低于对照组(P < 0.05)，视网膜干细胞移植组脑源性神经营养因子和胰岛素样生长因子Ⅰ蛋白的表达明显高于模型组(P < 0.05)，但低于对照组(P < 0.05)。 2.5 视网膜神经节细胞的凋亡情况由表3可以看出，模型组视网膜神经节细胞的凋亡数明显高于对照组(P < 0.05)，视网膜干细胞移植组的视网膜神经节细胞的凋亡数明显低于模型组(P < 0.05)，但高于对照组(P < 0.05)。 2.6 视网膜神经节细胞的计数由表4可以看出，模型组视网膜神经节细胞数明显低于对照组(P < 0.05)，视网膜干细胞移植组的视网膜神经节细胞数明显高于模型组(P < 0.05)，但低于对照组(P < 0.05)。"

References

[1] 林丁,陈琛.青光眼的视网膜神经节细胞损伤及其保护[J].中华眼科杂志,2005, 42(12):1144-1148.
[2] Kang MH,Yu DY.Distribution pattern of axonal cytoskeleton proteins in the human optic nerve head. Neural Regen Res. 2015; 10 (8): 1198-1200.
[3] Meyer JS, Katz ML, Maruniak JA, et al. Embryonic stem cell-derived neural progenitors incorporate into degenerating retina and enhance survival of host photoreceptors. Stem Cells. 2006;24(2):274-283.
[4] Bull ND, Limb GA, Martin KR. Human Müller stem cell (MIO-M1) transplantation in a rat model of glaucoma: survival, differentiation, and integration. Invest Ophthalmol Vis Sci. 2008;49(8):3449-3456.
[5] Wax MB, Tezel G, Kawase K, et al. Serum autoantibodies to heat shock proteins in glaucoma patients from Japan and the United States. Ophthalmology. 2001;108(2):296-302.
[6] Javier Carreras F.Pathogenesis of glaucoma: how to prevent ganglion cell from axonal destruction? . Neural Regen Res. 2014;9 (23): 2046-2047.
[7] Cheon EW, Park CH, Kang SS, et al. Betaxolol attenuates retinal ischemia/reperfusion damage in the rat. Neuroreport. 2003;14(15):1913-1917.
[8] Biermann J, Lagrèze WA, Dimitriu C, et al. Preconditioning with inhalative carbon monoxide protects rat retinal ganglion cells from ischemia/ reperfusion injury. Invest Ophthalmol Vis Sci. 2010;51(7):3784-3791.
[9] Vasudevan SK, Gupta V, Crowston JG. Neuroprotection in glaucoma. Indian J Ophthalmol. 2011;59 Suppl:S102-113.
[10] 柳浩然,杨长虹,高俊玮,等.神经干细胞移植对大鼠视神经损伤后节细胞的保护作用[J].中国微侵袭神经外科杂志, 2006,11(5):221-224.
[11] 项平,黄锦桃,李海标,等.骨髓间充质干细胞对成年大鼠视网膜节细胞再生的影响[J].解剖学杂志,2005,28(3): 252-254.
[12] Weiss JN, Levy S,Malkin A.Stem Cell Ophthalmology Treatment Study (SCOTS) for retinal and optic nerve diseases: a preliminary report. Neural Regen Res. 2015; 10 (6): 982-988.
[13] Yu HH,Cheng L,Cho KS.The potential of stem cell-based therapy for retinal repair. Neural Regen Res. 2014;9(11): 1100-1103.
[14] Zhou X, Xia XB, Xiong SQ. Neuro-protection of retinal stem cells transplantation combined with copolymer-1 immunization in a rat model of glaucoma. Mol Cell Neurosci. 2013;54:1-8.
[15] Lepski G, Maciaczyk J, Jannes CE, et al. Delayed functional maturation of human neuronal progenitor cells in vitro. Mol Cell Neurosci. 2011;47(1):36-44.
[16] Böhm MR, Pfrommer S, Chiwitt C, et al. Crystallin-β-b2-overexpressing NPCs support the survival of injured retinal ganglion cells and photoreceptors in rats. Invest Ophthalmol Vis Sci. 2012;53(13):8265-8279.
[17] Lawrence JM, Singhal S, Bhatia B, et al. MIO-M1 cells and similar muller glial cell lines derived from adult human retina exhibit neural stem cell characteristics. Stem Cells. 2007;25(8):2033-2043.
[18] Singhal S, Bhatia B, Jayaram H, et al. Human Müller glia with stem cell characteristics differentiate into retinal ganglion cell (RGC) precursors in vitro and partially restore RGC function in vivo following transplantation. Stem Cells Transl Med. 2012;1(3): 188-199.
[19] Johnson TV, Bull ND, Martin KR. Neurotrophic factor delivery as a protective treatment for glaucoma. Exp Eye Res. 2011;93(2):196-203.
[20] Voulgari-Kokota A, Fairless R, Karamita M, et al. Mesenchymal stem cells protect CNS neurons against glutamate excitotoxicity by inhibiting glutamate receptor expression and function. Exp Neurol. 2012; 236(1):161-170.
[21] Franquesa M, Hoogduijn MJ, Bestard O, et al. Immunomodulatory effect of mesenchymal stem cells on B cells. Front Immunol. 2012;3:212.
[22] Bull ND, Johnson TV, Welsapar G, et al. Use of an adult rat retinal explant model for screening of potential retinal ganglion cell neuroprotective therapies. Invest Ophthalmol Vis Sci. 2011;52(6): 3309-3320.
[23] Dalous J, Larghero J, Baud O. Transplantation of umbilical cord-derived mesenchymal stem cells as a novel strategy to protect the central nervous system: technical aspects, preclinical studies, and clinical perspectives. Pediatr Res. 2012;71(4 Pt 2):482- 490.
[24] Zhao T, Li Y, Tang L, et al. Protective effects of human umbilical cord blood stem cell intravitreal transplantation against optic nerve injury in rats. Graefes Arch Clin Exp Ophthalmol. 2011;249(7): 1021-1028.
[25] Chen M, Xiang Z, Cai J. The anti-apoptotic and neuro-protective effects of human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) on acute optic nerve injury is transient. Brain Res. 2013;1532:63-75.
[26] Jiang B, Zhang P, Zhou D, et al. Intravitreal transplantation of human umbilical cord blood stem cells protects rats from traumatic optic neuropathy. PLoS One. 2013;8(8):e69938.
[27] Robinton DA, Daley GQ. The promise of induced pluripotent stem cells in research and therapy. Nature. 2012;481(7381):295-305.
[28] Satarian L, Javan M, Kiani S, et al. Engrafted human induced pluripotent stem cell-derived anterior specified neural progenitors protect the rat crushed optic nerve. PLoS One. 2013;8(8):e71855.
[29] Wang T, Cong R, Yang H, et al. Neutralization of BDNF attenuates the in vitro protective effects of olfactory ensheathing cell-conditioned medium on scratch-insulted retinal ganglion cells. Cell Mol Neurobiol. 2011;31(3):357-364.
[30] Mead B, Logan A, Berry M, et al. Intravitreally transplanted dental pulp stem cells promote neuroprotection and axon regeneration of retinal ganglion cells after optic nerve injury. Invest Ophthalmol Vis Sci. 2013;54(12):7544-7556.
[31] Eiraku M, Takata N, Ishibashi H, et al. Self-organizing optic-cup morphogenesis in three-dimensional culture. Nature. 2011;472(7341):51-56.
[32] Hu Y, Ji J, Xia J, et al. An in vitro comparison study: the effects of fetal bovine serum concentration on retinal progenitor cell multipotentiality. Neurosci Lett. 2013;534:90-95.
[33] Chen G, Ma J, Shatos MA, et al. Application of human persistent fetal vasculature neural progenitors for transplantation in the inner retina. Cell Transplant. 2012;21(12):2621-2634.
[34] Parameswaran S, Balasubramanian S, Babai N, et al. Induced pluripotent stem cells generate both retinal ganglion cells and photoreceptors: therapeutic implications in degenerative changes in glaucoma and age-related macular degeneration. Stem Cells. 2010; 28(4):695-703.
[35] Chen M, Chen Q, Sun X, et al. Generation of retinal ganglion-like cells from reprogrammed mouse fibroblasts. Invest Ophthalmol Vis Sci. 2010;51(11): 5970-5978.
[36] Johnson TV, Bull ND, Martin KR. Identification of barriers to retinal engraftment of transplanted stem cells. Invest Ophthalmol Vis Sci. 2010;51(2):960-970.
[37] Singhal S, Lawrence JM, Salt TE, et al. Triamcinolone attenuates macrophage/microglia accumulation associated with NMDA-induced RGC death and facilitates survival of Müller stem cell grafts. Exp Eye Res. 2010;90(2):308-315.
[38] Kador KE, Goldberg JL. Scaffolds and stem cells: delivery of cell transplants for retinal degenerations. Expert Rev Ophthalmol. 2012;7(5):459-470.
[39] Cho JH, Mao CA, Klein WH. Adult mice transplanted with embryonic retinal progenitor cells: new approach for repairing damaged optic nerves. Mol Vis. 2012;18: 2658-2672.
[40] Reh TA, Levine EM. Multipotential stem cells and progenitors in the vertebrate retina. J Neurobiol. 1998; 36(2):206-220.
[41] Reh TA, Fischer AJ. Stem cells in the vertebrate retina. Brain Behav Evol. 2001;58(5):296-305.
[42] Ahmad I, Tang L, Pham H. Identification of neural progenitors in the adult mammalian eye. Biochem Biophys Res Commun. 2000;270(2):517-521.
[43] Tropepe V, Coles BL, Chiasson BJ, et al. Retinal stem cells in the adult mammalian eye. Science. 2000; 287(5460):2032-2036.
[44] Marquardt T, Gruss P. Generating neuronal diversity in the retina: one for nearly all. Trends Neurosci. 2002; 25(1):32-38.
[45] 俞海燕,吴文涛,王薇,等.成人骨髓间充质干细胞体外向视网膜细胞的诱导分化[J].中国药理学通报, 2014,30(6): 787-791.