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| 人牙周膜干细胞、脐带间充质干细胞对体外培养角膜缘干细胞的影响 | |
| 引用本文： | 王慧娴,高晓唯,蔡岩,李文静. 人牙周膜干细胞、脐带间充质干细胞对体外培养角膜缘干细胞的影响[J]. 眼科新进展, 2016, 0(11): 1015-1020. DOI: 10.13389/j.cnki.rao.2016.0271 |
| 作者姓名： | 王慧娴 高晓唯 蔡岩 李文静 |
| 作者单位： | 832003 新疆维吾尔自治区石河子市，石河子大学医学院（王慧娴，高晓唯）；830012 新疆维吾尔自治区乌鲁木齐市，解放军第474医院眼科中心（王慧娴，高晓唯，蔡岩，李文静） |
| 基金项目： | 新疆自治区科技支疆项目(编号:201491171)Project Plan of Science and Technology Assistance in Xinjiang Autonomous Region (201491171) |
| 摘 要： | 目的 比较人牙周膜干细胞（ｈｕｍａｎｐｅｒｉｏｄｏｎｔａｌｌｉｇａｍｅｎｔｓｔｅｍｃｅｌｌｓ，ＨＰＤＬＳＣｓ）、人脐带间充质干细胞（ｈｕｍａｎｕｍｂｉｌｉｃａｌｃｏｒｄｍｅｓｅｎｃｈｙｍａｌｓｔｅｍｃｅｌｌｓ，ＨＵＣＭＳＣｓ）作为饲养层培养角膜缘干细胞（ｌｉｍｂａｌｓｔｅｍｃｅｌｌｓ，ＬＳＣｓ）的优越性，从而筛选出扩增ＬＳＣｓ的最佳饲养层。方法 体外分离培养ＨＰＤＬＳＣｓ、ＨＵＣＭＳＣｓ，诱导细胞成脂、成骨分化，并检测细胞表面标志物的表达；将兔ＬＳＣｓ与ＨＰＤＬＳＣｓ、ＨＵＣＭＳＣｓ、ＮＩＨ-３Ｔ３共培养和无饲养层单独培养，比较各组克隆形成率，免疫荧光比较各组ＬＳＣｓ克隆团ＡＢＣＢ５、ＩＰＯ１３、ＣＫ３／１２的表达情况。结果 体外培养ＨＰＤＬＳＣｓ、ＨＵＣＭＳＣｓ均可向脂肪、成骨细胞分化，两种细胞均高表达间充质来源的表面标志ＣＤ９０，不表达ＣＤ４５；三组饲养层与兔ＬＳＣｓ共培养均可形成小克隆团， ＨＰＤＬＳＣｓ组、ＨＵＣＭＳＣｓ组、ＮＩＨ-３Ｔ３组、无饲养层组克隆形成率分别为（４．９０±０．９６）％、（４．１０±０．５６）％、（４．６７±０．７６）％、（０．８３±０．３５）％，四组间总体比较差异有统计学意义（Ｆ＝２２．０４７，Ｐ＝０．００）；ＨＰＤＬＳＣｓ组、ＨＵＣＭＳＣｓ组与ＮＩＨ-３Ｔ３组的克隆形成率差异均无统计学意义（均为Ｐ＞０．０５）；ＨＰＤＬＳＣｓ组、ＨＵＣＭＳＣｓ组、ＮＩＨ-３Ｔ３组与无饲养层组间差异均有统计学意义（均为Ｐ＜０．０１）。免疫荧光化学检测三种饲养层ＬＳＣｓ标志物ＡＢＣＢ５、ＩＰＯ１３呈高表达，ＣＫ３／１２呈低表达。结论 ＨＰＤＬＳＣｓ、ＨＵＣＭＳＣｓ均可作为替代饲养层用以培养ＬＳＣｓ。 |
| 关 键 词： | 人牙周膜干细胞 人脐带间充质干细胞 角膜缘干细胞 饲养层 |
Effects of periodontal ligament stem cells and human umbilical cord mesenchymal stem cells on microenvironment of corneal limbal stem cells cultured in vitro |
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| WANG Hui-Xian,GAO Xiao-Wei,CAI Yan,LI Wen-Jing. Effects of periodontal ligament stem cells and human umbilical cord mesenchymal stem cells on microenvironment of corneal limbal stem cells cultured in vitro[J]. Recent Advances in Ophthalmology, 2016, 0(11): 1015-1020. DOI: 10.13389/j.cnki.rao.2016.0271 | |
| Authors: | WANG Hui-Xian GAO Xiao-Wei CAI Yan LI Wen-Jing |
| Affiliation: | Shihezi University SchooL of Medicine( WANG Hui-Xian , GAO Xiao-Wei ) , Shihezi 832003 , Xinjiang Uygur Autonomous Region , China ; Ophthalmic Center , No. 474 Hospital of Chinese PLA ( WANG Hui-Xian , GAO Xiao-Wei, CAI Yan , LI Wen-Jing ) , Urumqi 830012 , Xinjiang Uygur Autonomous Region , China |
| Abstract: | Objective To find optimal feeding layer of amplification with limbal stem cells , compare the superiority of the human periodontal ligament stem cells ( HPDLSCs) and human umbilical cord mesenchymal stem cells ( HUCMSCs) as a breeding cultivation of corneal limbus stem cell layer , then find the more efficient feeder layer. Methods HPDLSCs and HUCMSCs were cultured in vitro .then the cells was induced and differentiated into fat. osteoblast cells . and the expression of cell surface markers were detected. The rabbit corneal limbal stem cells co-cultured with HPDLSCs . HUCMSCs.NIH-3T3 and no feeder layer, then each clone formation rate was counted, LSCs markers ABCB5 . IP013 . CK3/12 expression was compared by immunofluorescence between groups. Results HPDLSCs and HUCMSCs cultured in vitro could differentiate into fat,osteoblast cells and the two cells both expressed high surface of mesenchymal origin marks CD90 , not expressed CD45 . These groups could form cloning , the clone formation rate were (4. 90 +0. 96 ) % . (4. 10 + 0. 56) % , ( 4. 67 + 0. 76) % . ( 0. 83 + 0. 35 ) % in HPDLSCs ,HUCMSCs.NIH-3T3 and no feeder layer group , overall difference between the four groups was statistically sigruficant ( F= 22. 047 ,P < 0. 01) . There was no statistical significance of HPDLSCs.HUCMSCs groups compared with NIH-3T3 group ( all P > 0. 05 ) .but there were significance differences of HPDLSCs . HUCMSCs , NIH-3T3 groups compared with no feeder layer group ( all P < 0. 01) . Immunofluorescence showed three layer of LSCs markers ABCB5 ,IP013 were highly expressed, CK3/12 showed lower expression. Conclusion HPDLSCs. HUCMSCs both can be used as ideal substitute feeder layer to cultivate LSCs. |
| Keywords: | periodontal ligament stem cells umbilical cord mesenchymal stem cells corneal limbal stem cells feeder layer |
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