开元棋牌电脑 |本期目录/Table of Contents|

[1]彭昶璠,徐路星,罗世男,等.转化生长因子-β(TGF-β)空间动态分布的角膜基质创伤修复体外三维培养体系的构建[J].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2019,39(4):311-315.[doi:10.13389/j.cnki.rao.2019.0070]
 PENG Chang-Fan,XU Lu-Xing,LUO Shi-Nan,et al.Establishment of a three-dimensional culture system for corneal stromal wound healing in vitro with dynamic spatial distribution of TGF-β[J].Recent Advances in Ophthalmology,2019,39(4):311-315.[doi:10.13389/j.cnki.rao.2019.0070]
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转化生长因子-β(TGF-β)空间动态分布的角膜基质创伤修复体外三维培养体系的构建(/HTML)
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《开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷》[ISSN:1003-5141/CN:41-1105/R]

卷:
39卷
期数:
2019年4期
页码:
311-315
栏目:
实验研究
出版日期:
2019-04-05

文章信息/Info

Title:
Establishment of a three-dimensional culture system for corneal stromal wound healing in vitro with dynamic spatial distribution of TGF-β
作者:
彭昶璠徐路星罗世男王霜宁袁检宝李霞
530021 广西壮族自治区南宁市,广西医科大学第一附属医院眼科
Author(s):
PENG Chang-FanXU Lu-XingLUO Shi-NanWANG Shuang-NingYUAN Jian-BaoLI Xia
Department of Ophthalmology,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China
关键词:
角膜基质细胞转化生长因子-β三维培养创伤修复瘢痕化
Keywords:
corneal stromal cellstransforming growth factor-βthree-dimensional culturewound healingscarring
分类号:
R772
DOI:
10.13389/j.cnki.rao.2019.0070
文献标志码:
A
摘要:
目的 研究构建模拟角膜基质创伤修复过程中转化生长因子-β(transforming growth factor-β,TGF-β)空间动态分布的体外三维培养系统。方法 从新鲜牛眼中分离角膜基质细胞,用含体积分数10%胎牛血清(fetal bovine serum,FBS)的DMEM/F12培养基进行培养,而后构建Pellet体外三维培养模型进行培养。将Pellet分为有透析袋组和无透析袋组,置入Transwell小室系统上下室中培养48 h后换液,上室为0.50 μg·L-1TGF-β1+0.25 μg·L-1TGF-β2+体积分数10%FBS,下室为体积分数10%FBS,分别于培养后72 h观察Pellet培养模型的形态变化并采用Real-time PCR法分别检测2组上下室Pellet中α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、纤维连接蛋白(fibronectin,FN)、Ⅰ型胶原(collagen Ⅰ,Col Ⅰ)和Col Ⅲ mRNA相对表达量。结果 培养后72 h Pellet均呈团状生长。有透析袋组Transwell小室系统上、下室α-SMA mRNA表达量分别为1.595±0.025、1.148±0.009,FN mRNA表达量分别为1.090±0.011、0.844±0.015,Col Ⅰ mRNA表达量分别为1.445±0.035、1.165±0.008,Col Ⅲ mRNA表达量分别为1.726±0.031、1.314±0.020,Col Ⅲ/Col Ⅰ比值分别为1.126±0.019、0.957±0.013,上下室各项指标比较差异均有统计学意义(均为P=0.000)。无透析袋组Transwell小室系统上、下室α-SMA mRNA表达量分别为1.363±0.018、1.360±0.002,FN mRNA表达量分别为0.946±0.017、0.952±0.012,Col Ⅰ mRNA表达量分别为1.261±0.011、1.258±0.029,Col Ⅲ mRNA表达量分别为1.459±0.027、1.462±0.033,Col Ⅲ/Col Ⅰ比值分别为1.157±0.029、1.163±0.090,上下室各项指标比较,差异均无统计学意义(均为P>0.05)。无透析袋组上室细胞中α-SMA、FN、Col Ⅰ和Col Ⅲ mRNA的相对表达量与有透析袋组上室、有透析袋组下室比较,差异均有统计学意义(均为P<0.05)。无透析袋组下室细胞中α-SMA、FN、Col Ⅰ和Col Ⅲ mRNA的相对表达量与有透析袋组上室、有透析袋组下室比较,差异均有统计学意义(均为P<0.05)。结论 Transwell小室系统与透析袋相结合可构建模拟TGF-β空间动态分布的体外三维培养系统。
Abstract:
Objective To establish a three-dimensional culture system simulating the spatial dynamic distribution of transforming growth factor-β (TGF- β) during corneal matrix healing in vitro.Methods Corneal stromal cells were isolated from fresh bovine eyes and cultured in DMEM/F12 culture medium containing volume fraction of 10%FBS.Then a three-dimensional culture model of Pellet was established for culture.Pellets were divided into dialysis tubes group and non-dialysis tubes group.After 48 h of culture in the upper and lower compartments of Transwell chamber system,the fluid in the upper chamber was changed to 0.50 μg·L-1 TGF-β1+0.25 μg·L-1 TGF-β2+volume fraction of 10% FBS,and volume fraction of 10%FBS in the lower chambers.The morphological changes of Pellets’ culture model were observed 72 h after culture.Real-time PCR was used to detect the relative expressions of α-smooth muscle actin (α-SMA),fibronectin (FN),type Ⅰ collagen (Col Ⅰ) and type Ⅲ collagen (Col Ⅲ) mRNA in each group.Results After 72 hours of culture,Pellets grew into clusters.In the upper and lower chambers of the dialysis tubes group,the expression level of α-SMA mRNA was 1.595±0.025 and 1.148±0.009,respectively;FN mRNA was 1.090±0.011 and 0.844±0.015,respectively;Col I mRNA was 1.445±0.035 and 1.165±0.008,respectively;Col Ⅲ mRNA was 1.726±0.031 and 1.314±0.020,respectively;and the ratio of Col Ⅲ/Col Ⅰ was 1.126±0.019 and 0.957±0.013,respectively.The differences between the upper and lower chambers were statistically significant (all P=0.000).In the upper and lower chambers of the non-dialysis tubes group,the expression level of α-SMA mRNA was 1.363±0.018 and 1.360±0.002,respectively;FN mRNA was 0.946±0.017 and 0.952±0.012,respectively;Col Ⅰ mRNA expression was 1.261±0.011 and 1.258±0.029,respectively;Col Ⅲ mRNA was 1.459±0.027 and 1.462±0.033,respectively;and the ratio of Col Ⅲ/Col Ⅰ was 1.157±0.029 and 1.163±0.090,respectively.There was no significant statistical difference between upper and lower chambers of non-dialysis tubes group (all P>0.05).The relative expression levels of α-SMA,FN,Col Ⅰ and Col Ⅲ mRNA in the upper chamber of the non-dialysis tubes group were significantly different from those of the upper and lower chambers of the dialysis tubes group (all P<0.05).The relative expressions of α-SMA,FN,Col Ⅰ and Col Ⅲ in the lower chamber of the non-dialysis tubes group were also significantly different from those of the upper and lower chamber of the dialysis tubes group (all P<0.05).Conclusion The combination of the Transwell chamber system and dialysis tubes is able to establish a three-dimensional culture system which can simulate the spatial dynamic distribution of TGF-β in vitro.

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备注/Memo

备注/Memo:
国家自然科学基金资助(编号:81360144、81060076);广西自然科学基金资助(编号:2017-GXNSFAA198250)
更新日期/Last Update: 2019-04-15