开元棋牌电脑 |本期目录/Table of Contents|

[1]周瑛,孙秋萍,蔡岩,等.3D共培养模型对体外培养兔角膜缘干细胞的影响[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):331-335.[doi:10.13389/j.cnki.rao.2019.0074]
 ZHOU Ying,SUN Qiu-Ping,CAI Yan,et al.Effect of three-dimensional co-culture model on rabbit corneal limbal stem cells culture in vitro[J].Recent Advances in Ophthalmology,2019,39(4):331-335.[doi:10.13389/j.cnki.rao.2019.0074]
点击复制

3D共培养模型对体外培养兔角膜缘干细胞的影响(/HTML)
分享到:

《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:
331-335
栏目:
实验研究
出版日期:
2019-04-05

文章信息/Info

Title:
Effect of three-dimensional co-culture model on rabbit corneal limbal stem cells culture in vitro
作者:
周瑛孙秋萍蔡岩高晓唯
830000 新疆维吾尔自治区乌鲁木齐市,新疆医科大学(周瑛,高晓唯);830000 新疆维吾尔自治区乌鲁木齐市,解放军第四七四医院眼科(周瑛,孙秋萍,蔡岩,高晓唯)
Author(s):
ZHOU YingSUN Qiu-PingCAI YanGAO Xiao-Wei
Xinjiang Medical University(ZHOU Ying,GAO Xiao-Wei),Urumqi 830000,Xinjiang Uygur Autonomous Region,China;Ophthalmic Center,No.474 Hospital of Chinese PLA(ZHOU Ying,SUN Qiu-Ping,CAI Yan,GAO Xiao-Wei),Urumqi 830000,Xinjiang Uygur Autonomous Region,China
关键词:
角膜缘干细胞纤维蛋白胶羊膜3D共培养
Keywords:
limbal stem cellsfibin glueamniotic membrane3D co-culture
分类号:
R772.2
DOI:
10.13389/j.cnki.rao.2019.0074
文献标志码:
A
摘要:
目的 探讨三维(three-dimension,3D)共培养模型对体外扩增兔角膜缘干细胞(limbal stem cells,LSCs)的作用。方法 体外培养兔LSCs,以牙周膜干细胞(human periodontal stem cell,HPDLSCs)作为饲细胞建立共培养模型,根据载体不同分为3组,纤维蛋白胶组:将兔LSCs、HPDLSCs、纤维蛋白胶3D共培养;羊膜组:将兔LSCs、HPDLSCs、羊膜3D共培养;对照组:将兔LSCs、HPDLSCs共培养。共培养5 d后,用倒置显微镜、HE染色、扫描电镜观察兔LSCs的细胞形态,采用免疫荧光染色检测各组兔LSCs阳性标志物p63的表达,并比较各组p63的阳性率。结果 共培养5 d后,倒置显微镜、HE染色观察细胞形态,羊膜组、纤维蛋白胶组细胞排列紧密,呈大小不等的团块状集落,细胞大小基本一致;对照组细胞排列相对稀疏;扫描电镜下观察见纤维蛋白胶组和羊膜组细胞与载体贴附良好。免疫荧光染色显示纤维蛋白胶组、羊膜组和对照组LSCs的p63阳性率分别为(69.93±8.76)%、(78.36±8.56)%和(58.59±8.31)%,三组间总体比较差异有统计学意义(F=9.43,P=0.000)。两两比较中纤维蛋白胶组、羊膜组LSCs的p63阳性率均大于对照组,差异均有统计学意义(均为P<0.05)。纤维蛋白胶组、羊膜组的p63阳性率比较,差异无统计学意义(P>0.05)。结论 以纤维蛋白胶、羊膜作为载体,HPDLSCs作为饲细胞的3D共培养模型有助于体外扩增兔LSCs。
Abstract:
Objective To investigate the effect of three-dimensional(3D) co-culture model on the expansion of rabbit limbal stem cells in vitro.Methods The rabbit limbal stem cells (LSCs) were cultured in vitro,and the human periodontal stem cells (HPDLSCs) were served as the feeder cells to establish of co-culture model.According to different carriers,the cells were divided into three groups:the fibrin glue group,in which LSCs were co-cultured with fibrin glue and HPDLSCs;the amniotic membrane group,in which LSCs were co-cultured with amniotic membrane and HPDLSCs;and the control group,in which LSCs were co-cultured only with the HPDLSCs.After co-culture for 5 days,we used inverted microscope,hematoxylin and eosin staining and scanning electron microscopy to observe the cells morphology,and then used the immunofluorescence staining to detect the expression of the LSCs positive marker p63,followed by the comparison of the positive rate of p63 in each group.Results After co-cultured for five days,the results from inverted microscope and hematoxylin and eosin staining showed that cells in the fibrin glue group and the amniotic membrane group were uniform and serried arrangement,and presented cluster-like colonies with varying sizes,and cell sizes were uniformly,whereas cells in the control group were relatively sparse.Scanning electron microscopy showed that the cells were well adhered to the carrier in the fibrin glue group and the amniotic membrane group.Immunofluorescence staining showed that the positive rates of p63 in the fibrin glue group,the amniotic membrane group,the control group were (69.93±8.76)%,(78.36±8.56)%,(58.59±8.31)%,respectively,and there was significant difference among the three groups (F=9.43,P=0.000).In pairwise comparison,the positive rates of p63 in the fibrin glue group and amniotic membrane group were both higher than those of the control group,and there was significant differences (both P<0.05),but there was no significant difference in the positive rate of p63 between the fibrin glue group and amniotic membrane group (P>"0.05).Conclusion The co-culture 3D model is helpful for in vivo expansion of rabbit LSCs.

参考文献/References:

[1] PELLEGRINI G,RAMA P,DI R A,PANARAS A,DE L M.Concise review:hurdles in a successful example of limbal stem cell-based regenerative medicine[J].Stem Cells,2014,32(1):26-34.
[2] SCHL TZER-SCHREHARDT U,KRUSE F E.Identification and characterization of limbal stem cells[J].Exp Eye Res,2005,81(3):247-264.
[3] XU B,FAN T J,ZHAO J,SUN A,WANG R X,HU X Z,et al.Transplantation of tissue-engineered human corneal epithelium in limbal stem cell deficiency rabbit models[J].Int J Ophthalmol,2012,5(4):424-429.
[4] HOSSEINKHANI M,MEHRABANI D,KARIMFAR M H,BAKHTIYARI S,MANAFI A,SHIRAZI R.Tissue engineered scaffolds in regenerative medicine[J].World J Plast Surg,2014,3(1):3-7.
[5] LI Y,GU Z Y,LV Q F,YOU J B,LIU F F.Primary culture of human periodontal ligament stem cells and dental pulp stem cells by tissue enzymatic digestion method[J].J Clin Rehabil Tis Eng Res,2012,16(32):5993-5998.
李颖,谷子芽,吕秋峰,尤金彪,刘芳菲.酶解组织块法原代培养人牙周膜干细胞和牙髓干细胞[J].中国组织工程研究,2012,16(32):5993-5998.
[6] HU R,GAO J,LIANG Y R,HUANG Y,LI H,SU M.An experimental study of cultivating rat limbal stem cells in vitro on amniotic membrane[J].Chongqing Med,2016,45(23):3233-3236.
胡蓉,高杰,梁雅茹,黄悦,李红,苏敏.以羊膜为载体体外培养大鼠角膜缘干细胞的实验研究[J].重庆医学,2016,45(23):3233-3236.
[7] ZHOU Y.Effects of feeder layer cell membrane fluidity on induced pluripotent stem cells (miPS) in mice[D].Chang chun:Jilin University,2014.
周悦.饲养层细胞膜流动性对小鼠诱导性多能干细胞(miPS)的影响 [D].长春:吉林大学,2014.
[8] WANG H X,GAO X W,CAI Y,LI W J.Effects of periodontal ligament stem cells and human umbilical cord mesenchymal stem cells on microenvironment of corneal limbal stem cells cultured in vitro[J].Rec Adv Ophthal,2016,36(11):1015-1020.
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].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2016,36(11):1015-1020.
[9] ZHANG X,SUN H,LI X,YUAN X,ZHANG L,ZHAO S.Utilization of human limbal mesenchymal cells as feeder layers for human limbal stem cells cultured on amniotic membrane[J].J Tissue Eng Reg Med,2010,4(1):38-44.
[10] MEI H,GONZ LEZ S,NAKATSU M N,BACLAGON E R,CHEN F V,DENG S X.Human adipose-derived stem cells support the growth of limbal stem/progenitor cells[J].PLoS One,2017,12(10):225-240.
[11] ZHANG W,FANG M,BAO D Y,DING X C,TONG X,QIN H Y.Effects of DAPT on the proliferation and osteogenic differentiation of human periodontal ligment stem cells[J].Oral Biomed,2018,9(1):7-11.
张文,方明,鲍东昱,丁晓晨,童昕,秦海燕.DAPT对牙周膜干细胞体外增殖及成骨分化的影响[J].口腔生物医学,2018,9(1):7-11.
[12] SAICHANMA S,BUNYARATVEJ A,SILA-ASNA M.In vitro transdifferentiation of corneal epithelial-like cells from human skin-derived precursor cells[J].Int J Ophthalmol,2012,5(2):158-163.
[13] BOURNE G L.The microscopic anatomy of the human amnion and chorion[J].Am J Obstet Gynecol,1960,79(6):1070-1073.
[14] UTHEIM T P,AASS UTHEIM ,SALVANOS P,JACKSON C J,SCHRADER S,GEERLING G,et al.Concise review:altered versus unaltered amniotic membrane as a substrate for limbal epithelial cells[J].Stem Cells Transl Med,2018,7(5):415-427.
[15] MOHAN R,BAJAJ A,GUNDAPPA M.Human amnion membrane:potential applications in oral and periodontal field[J].J Int Soc Pre Comm Dent,2017,7(1):15-21.
[16] NOORI A,ASHRAFI S J,VAEZ-GHAEMI R,HATAMIAN-ZAREMI A,WEBSTER T J.A review of fibrin and fibrin composites for bone tissue engineering[J].Int J Nanomed,2017,12:4937-4961.
[17] BROWN A C,BARKER T H.Fibrin-based biomaterials:Modulation of macroscopic properties through rational design at the molecular level[J].Acta Biomater,2014,10(4):1502-1514.
[18] LINSLEY C S,WU B M,TAWIL B.Mesenchymal stem cell growth on and mechanical properties of fibrin-based biomimetic bone scaffolds[J].J Biomed Mater Res,2016,104(12):2945-2953.
[19] MEHRZAD B,MILAD M,AZAM S,NAVID N,MANICA N,FATEMEH M,et al.Optimization of three dimensional culturing of the HepG2 cell line in fibrin scaffold[J].Hepat Mon,2015,15(3):e22731.
[20] XIONG L J,ZHENG T,HU Y Z.Co-culture of mouse embryonic fibroblasts and fibrin with corneal limbal stem cells for viability and phenotype maintenance[J].Acta Med Univ Sci Technol Huazhong,2012,41(6):680-683.
熊林杰,郑恬,胡义珍.鼠角膜缘干细胞3D共培养模型的建立及生存率、表型维持[J].华中科技大学学报(医学版),2012,41(6):680-683.

相似文献/References:

[1]李翠霞 洪晶 曲洪强.体外培养不同代角膜缘干细胞特性分析[J].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2012,32(7):.
[2]张伟 杨为中 金敏 高宗银 陈晓燕 许艳丽 张玉平.一种新的角膜缘干细胞分选方法[J].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2012,32(2):.
[3]黄 焱 杨 静 张 娣 马惠杰 廖 凯沈 平 袁 安.自体角膜缘干细胞移植与常规翼状胬肉切除术疗效比较[J].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2009,29(1):.
[4]李芳,李轩.间充质干细胞在角膜重建中的应用研究进展[J].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2015,35(11):1083.[doi:10.13389/j.cnki.rao.2015.0297]
 LI Fang,LI Xuan.Research progress on applications of MSC in cornea reconstruction[J].Recent Advances in Ophthalmology,2015,35(4):1083.[doi:10.13389/j.cnki.rao.2015.0297]
[5]王秀,杨军,宁玉贤,等.纤维蛋白胶在翼状胬肉手术中应用的研究进展[J].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2016,36(2):186.[doi:10.13389/j.cnki.rao.2016.0051]
 WANG Xiu,YANG Jun,NING Yu-Xian,et al.Recent advances in fibrin glue applied in pterygium surgery[J].Recent Advances in Ophthalmology,2016,36(4):186.[doi:10.13389/j.cnki.rao.2016.0051]
[6]杜娟,姜楠,邵毅,等.纤维蛋白胶治疗角膜板栗刺伤的临床研究[J].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2016,36(4):336.[doi:10.13389/j.cnki.rao.2016.0091]
 DU Juan,JIANG Nan,SHAO Yi,et al.Application of fibrin glue on corneal chestnut thorns injury[J].Recent Advances in Ophthalmology,2016,36(4):336.[doi:10.13389/j.cnki.rao.2016.0091]
[7]王惠学,范先群.干细胞与角膜眼表重建研究进展[J].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2016,36(8):788.[doi:10.13389/j.cnki.rao.2016.0211]
 WANG Hui-Xue,FAN Xian-Qun.Research progress on stem cells and corneal ocular surface reconstruction[J].Recent Advances in Ophthalmology,2016,36(4):788.[doi:10.13389/j.cnki.rao.2016.0211]
[8]王慧娴,高晓唯,蔡岩,等.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].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2016,36(11):1015.[doi:10.13389/j.cnki.rao.2016.0271]
 WANG Hui-Xian,GAO Xiao-Wei,CAI Yan,et al.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,2016,36(4):1015.[doi:10.13389/j.cnki.rao.2016.0271]
[9]马文贝,冯松福,韦诗羽,等.高糖抑制角膜缘干细胞间质性及其迁移能力的研究[J].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2018,38(5):407.[doi:10.13389/j.cnki.rao.2018.0095]
 MA Wen-Bei,FENG Song-Fu,WEI Shi-Yu,et al.High glucose suppresses the corneal limbal stem cell migration and changes the cell phenotype[J].Recent Advances in Ophthalmology,2018,38(4):407.[doi:10.13389/j.cnki.rao.2018.0095]
[10]赵丹丹,杨阳,陈景尧,等.封闭结膜和Tenon囊间隙联合羊膜移植及纤维蛋白胶治疗睑球粘连的疗效观察[J].开元棋牌电脑_开元棋牌一直在维护_开元棋牌怎么对刷,2018,38(10):964.[doi:10.13389/j.cnki.rao.2018.0228]
 ZHAO Dan-Dan,YANG Yang,CHEN Jing-Yao,et al.Efficacy observation of sealing the gap between conjunctiva and Tenon capsule combined with amniotic membrane transplantation and fibrin glue in the treatment of symblepharon[J].Recent Advances in Ophthalmology,2018,38(4):964.[doi:10.13389/j.cnki.rao.2018.0228]

备注/Memo

备注/Memo:
新疆维吾尔自治区科技支疆项目(编号:201491171);解放军第四七四医院重点扶持科研项目(编号:2017474006、2018474008)
更新日期/Last Update: 2019-04-15