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...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.展开更多
文摘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.
