胎盘间充质干细胞对角膜缘干细胞干性维持及扩增能力影响  

The impact of human placental mesenchymal stem cells on the ability to maintain and amplify the limbal stem cells

作　　者：王慧娴[1,2] 高晓唯[1,2] 杨于力[3] 李沂键 谢汉平[3] 蔡岩[2] 李文静[2] Wang Huixian Gao Xiaowei Yang Yuli Li Yijian Xie Hanping Cai Yan Li Wenjing(Shihezi University School of Medicine, Shihezi 832003, China Ophthalmic Center of the No.474 Hospital of Chinese PLA, Urumqi 830012, China Southwest Hospital Affiliated the Third Military Medical University, Chongqing 400038, China)

机构地区：[1]石河子大学医学院,新疆832003 [2]解放军第474医院眼科中心,乌鲁木齐830012 [3]第三军医大学附属西南医院,重庆400038

出　　处：《中国实用眼科杂志》2017年第1期84-88,共5页Chinese Journal of Practical Ophthalmology

基　　金：新疆维吾尔自治区科技支疆项目计划（201491171）

摘　　要：目的探讨人源性胎盘间充质干细胞（HPMSCs）能否用于体外扩增角膜缘干细胞（LSCs）并维持其干细胞特性。方法实验研究。对2015年10月至2016年6月在新疆眼科大学临床学院行体外分离培养HPMSCs并行成脂、成骨和细胞表面标志鉴定；兔LSCs与HPMSCs、NIH-3T3共培养并计算比较各组克隆形成率（CFE）；免疫荧光检测两组LSCs克隆团干细胞特性表达情况。结果体外培养HPMSCs具有成脂和成骨潜能；高表达CD29、CD44，不表达CD34、CD45、HLA-DR；与兔LSCs共培养8天后，两种饲养组均可形成较大LSCs克隆团，HPMSCs、NIH-3T3饲养组以及无饲养层组LSCs的CFE为（3．02±0．73）％、（4．11±0．69）％、（0．24±0．08）％，三组间总体差异有统计学意义（F=34．87，P〈0．01）；HPMSCs饲养组与NIH-3T3饲养组的CFE差异无统计学意义（t=2．27，P〉O．05）；HPMSCs、NIH-3T3饲养组与无饲养细胞组问均差异有统计学意义（t=5．82、8．10，P〈0．01）。免疫荧光化学检测两种饲养层LSCs克隆团标志物，IP013均呈高表达，CK3／12呈低表达，各饲养组干细胞标志物表达情况无显著差异。结论HPMSCs能扩增LSCs并维持其干细胞特性，可作为替代3T3的理想饲养层。Objective To look for maintaining limbal stem cells （LSCs） unditlerentiated growth of human-derived cells as the feeder layer and investigating placenta derived mesenchymal stem cells （HPMSCs） can be used in vitro amplification LSCs and maintain their stem cell properties. Meth- ods In vitro isolation and culture HPMSCs, induce adipogenic, osteogenic differentiation, flow cytom- etry cell surface markers. Rabbits＇ LSCs respectively co-cultured with HPMSCs, NIH-3T3 to calcu- late and compare the group cloning efficiency （CFE）. Immunofluorescence examination can detect that if IPO13, CK3/12 existed. Results In vitro culture HPMSCs had the potency to differentiate in- to adipocytes and osteoblasts. They had strong proliferation ability and showed positive expression of CD29, CD44 and negative of CD34, CD45, HLA-DR. After a total of 8 days of co-culturing rab- bits＇ LSCs, the two feeder layers can form a larger LSCs clone group. The LSCs CFE of HPM- SCs, NIH-3T3 feeder and feeder-free group was （3.02±0.73）%, （4.11±0.69）%, （0.24±0.08）%, respec-tively. The overall difference among the three groups was statistically significant （F =34.87, P 〈 0.01）. HPMSCs feeding group and NIH-3T3 feeder group had no statistically significant difference （t =2.27, P 〉0.05）; Among HPMSCs, NIH-3T3 feeder group and cell-free group were different （t = 5.82, 8.10, P 〈0.01）. Immunofluorescence detection of two feeder layers LSCs clone group markers IPO13 showed high expression, CK3/12 showed low expression, each group had no significant differ- ences in the expression of stem cell markers. Conclusions HPMSCs can amplify LSCs and main- tain their stem characteristics, which can be used as an ideal substitute of 3T3 feeder layer.

关 键 词：胎盘间充质干细胞 饲养层 共培养 

分 类 号：R394.2[医药卫生—医学遗传学]