胎儿角膜缘干细胞的培养和生物学特性观察

角膜缘干细胞(limbalstemcells,LSC)及利用角膜缘干细胞移植(limbalstemcellstransplantation,LSCT)重建眼表的研究正在国内外广泛展开。而实行LSCT其前提是必须有LSC供体,尽管当前LSC的来源有自体、同种异体和体外培养的自体及同种异体等几种,但均有其局限性。人胎儿细胞的增殖和分化潜能较成人同类细胞大,此外,胎儿的免疫系统没有发育完善,胎儿组织存在免疫宽容现象[1]。贾卉等[2]研究还发现,胎儿角膜中与免疫反应有关的细胞和细胞间黏附分子较成人少,故胎儿器官、组织或细胞移植较少甚至无免疫排斥反应发生。因此,为探寻一种新的…

Ex vivo culture and biological characteristics observation of fetal limbal stem cells

ObjectiveTo investigate the culture and identification of fetal limbal stem cells ex vivo.MethodsFetal limbal epithelial tissue was cultured in vitro by enzymatic digestion method and the morphological characteristics of cultured cells were recorded.The fetal limbus and cultured cells were detected by enzyme immunocytochemistry staining with a series of monoclonal antibodies.The surface characteristics of primary culture cells were observed by scanning electron microscope.ResultsFetal limbal epithelial cells grew swiftly in primary culture and kept high proliferating rate in subculture.Many cells from basal layer of fetal limbus were absent staining for AE5 and positive for AE1,PCNA and HLA-DR,however,majority of primary culture cells were negative for AE5 and HLA-DR and positive for AE1 and PCNA,and most cells in the third passage were AE5 positive.The morphological characteristics of primary culture cells were mainly spherical shape or short column shape with plenty of processes and microvilli on the surface under the scanning electron microscope.ConclusionFetal limbal stem cells with high proliferating ability and low antigenicity can be successfully cultured by enzymatic digestion method,showing a broad applying prospect in ocular surface reconstruction.