不同细胞外基质对人胚胎干细胞体外定向诱导分化为角化细胞的影响  

Influence of different extracellular matrix to induced differentiation of Human Embryonic Stem Cells to Keratinocytes in vitro

作　　者：赵厚明[1,2] 李晗卿[1,2] 周海文[1,2] 

机构地区：[1]上海交通大学医学院 [2]上海交通大学医学院附属第九人民医院口腔黏膜科.上海市口腔医学重点实验室,上海200011

出　　处：《临床口腔医学杂志》2015年第3期148-151,共4页Journal of Clinical Stomatology

基　　金：国家自然科学基金项目资助(81170968);2013~2014年度国家临床重点专科建设项目资助(国卫办医函[2013]544号)

摘　　要：目的:探讨不同细胞外基质对胚胎干细胞源性角化细胞生长、增殖及分化的影响。方法:体外培养的人类胚胎干细胞经RA、BMP4、AA诱导后,分别接种到以Matrigel(A组)、Ⅰ型胶原(B组)、Ⅳ型胶原(C组)、明胶(D组)包被的培养皿中培养,E组为无包被培养皿作为空白对照组。通过倒置显微镜观察细胞生长情况;用CCK8法检测细胞的增殖能力;并通过Western Blot检测CK14表达量,测定各组灰度值。结果:A组生长的E-KCs细胞最快到达生长对数期,增殖能力最强,且细胞集落形成率最高,与其他组比较,差异具有统计学意义(P<0.05);A组与正常口腔黏膜组(阳性对照)CK14表达量条带灰度值差异无统计学意义,但与B、C、D组比较差异具有统计学意义(P<0.01)。结论:Matrigel与其他3种细胞外基质相比能显著提高人类胚胎干细胞向角化细胞诱导分化的能力,并能促进E-KCs生长和增殖。Objective：To research different extracellular matrix＇s influence to kerotinocyte derived from Human Em-bryonic Stem Cells about growth,proliferation and differentiation. Method：Culture the human embryonic stem cells In vitro and induced by RA、BMP4、AA,then culture the cells in the dishs coated with Matrigel （A）、CollagenⅠ（B）、CollagenⅣ（C）、Gelatin（D）,and nothing coating the dish（E）. TO observe the cell growth by inverted microscope,Detect cell prolifer-ation With CCK8,The expression of CK14 is detected by the western-blotting. Result：The E-KCs grow faster and have the stronger proliferation abilities than other groups. Cell colony formation rate is high and T test shows P 〈0.05 compared with other groups. Western-blotting shows that the expression of CK14 of A group is higher than other groups ,P〈0.01,the differ-ence is statistically significant. Conclusion：Matrigel can significantly improve the ability of human embryonic stem cells in-ducing into the keratinocytes, and can promote the proliferation of E-KCs.

关 键 词：胚胎干细胞 细胞外基质 诱导分化 角化细胞 

分 类 号：Q813.1[生物学—生物工程]