Chondroitinase with adipose-derived mesenchymal stem cell transplantation: a therapeutic strategy for retinal degeneration in rats

doi:10.3969/j.issn.2095-4344.2017.09.014

Abstract

Abstract:

BACKGROUND: Studies have found that chondroitin sulfate proteoglycans degradation with chondroitinase can promote the migration of Müller cells on the retina, but whether it could promote the migration of adipose-derived mesenchymal stem cells in retinal degeneration rats is unclear.
OBJECTIVE: To investigate the effect of chondroitin sulfate proteoglycans degradation with chondroitinase on adipose-derived mesenchymal stem cell treatment for retinal degeneration in rats.
METHODS: Human adipose-derived mesenchymal stem cells were isolated and cultured. A retinal degeneration rat model was established followed by administration of adipose-derived mesenchymal stem cells+chondroitinase into the subretinal space. The migration of adipose-derived mesenchymal stem cells and retinal cell apoptosis in rats after transplantation were observed.
RESULTS AND CONCLUSION: The human adipose-derived mesenchymal stem cells could be successfully cultured. The labeling rate of BrdU to the human adipose-derived mesenchymal stem cells was more than 90.0%. At 7 days after modeling, the outer nuclear layer of the retina was collapsed, and a large amount of photoreceptor cells were dissected. The outer nuclear layer was attached to the Bruch’s membrane, and the retina was arched. The central retina and peripheral retina were damaged. In normal rats, the retinal layers were clear, and the photoreceptor cells arranged regularly; and the retinal pigment epithelium was complete. The migration rate of adipose-derived mesenchymal stem cells in adipose-derived mesenchymal stem cells+chondroitinase group was higher than that in adipose-derived mesenchymal stem cells group (P < 0.05). There was no significant difference in the apoptotic rate of retinal cells between the two groups (P > 0.05). These experimental findings show that chondroitin sulfate proteoglycans degradation with chondroitinase can enhance the migration ability of human adipose-derived mesenchymal stem cells on the retina.

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: Retinal Degeneration, Cell Movement, Tissue Engineering

CLC Number:

R394.2

Chu Miao, Ju Chao-juan, Zhao Li-na, Cui Zeng-min, Song Geng. Chondroitinase with adipose-derived mesenchymal stem cell transplantation: a therapeutic strategy for retinal degeneration in rats[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(9): 1384-1389.

Figures/Tables 2

2.1 人脂肪间充质干细胞培养脂肪间充质干细胞原代培养一两天时少量多角细胞或梭形细胞，3-5 d后梭形细胞增多，7-10 d时细胞形成集落，呈团簇状生长，12-14 d时细胞达80%以上融合(图1)。经传代培养，多角形细胞不断减少，传代至第3代时，多角形细胞基本消失，均为梭形细胞。 2.2 人脂肪间充质干细胞鉴定结果第3代人脂肪间充质干细胞CD29、CD44、CD90、CD105的阳性表达率分别为99.31%、98.42%、99.53%和98.02%，C34、CD45的阳性表达率为0.19%和0.62%。表明培养细胞为脂肪间充质干细胞。 2.3 人脂肪间充质干细胞标记效果荧光显微镜下可见，经BrdU标记的人脂肪间充质干细胞的细胞核呈红色荧光，标记率达90.0%以上(图2)。 2.4 实验动物数量分析动物实验进入结果分析的实验动物数量为24只，中间无脱落。 2.5 大鼠视网膜变性模型成功建立建模后7 d，视网膜外核层塌陷，光感受器细胞大量外节迸解，外核层贴附在Bruch’s膜上，视网膜呈拱桥样，中央视网膜和外周视网膜均受到损伤。正常大鼠视网膜各层清晰，光感受器细胞排列规律，视网膜色素上皮层完整(图3)。 2.6 脂肪间充质干细胞迁移率脂肪间充质干细胞移植组移植后脂肪间充质干细胞主要分布在视网膜下腔，只有少量脂肪间充质干细胞迁移到视网膜外核层以内；脂肪间充质干细胞+硫酸软骨素酶组脂肪间充质干细胞大部分迁移到视网膜外核层内，脂肪间充质干细胞+硫酸软骨素酶组脂肪间充质干细胞迁移率高于脂肪间充质干细胞组(P < 0.05；图4，表1)。 2.7 视网膜细胞凋亡情况各组视网膜细胞凋亡以外核层为主。脂肪间充质干细胞组和脂肪间充质干细胞+硫酸软骨素酶组视网膜细胞凋亡率均低于对照组(P < 0.05)，脂肪间充质干细胞组和脂肪间充质干细胞+硫酸软骨素酶组视网膜细胞凋亡率比较差异无显著性意义(P > 0.05；图5，表2)。"

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