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Using bovine pituitary extract to increase proliferation of keratocytes and maintain their phenotype in vitro

International Journal of Ophthalmology(English edition) 2013年第6期6卷 758-765页

作者： Zhong-Zhong Xu Zhi-Jie Li Lian-Xin Du Jing Li Li-Ya Wang Department of Ophthalmology the First Affiliated Hospital of Zhengzhou University Key Laboratory of Corneal Disease Henan Eye Institute Department of Pediatrics Baylor College of Medicine

AIM:To investigate the effects of bovine pituitary extract on the proliferation of keratocytes and maintaining the keratocyte phenotype/n v/***:Single keratocytes were isolated by enzyme digestion for in vitro *** groups were designed according to the different culture media:a bovine pituitary extract(BPE)group,a fetal bovine serum(FBS)group and the control *** phenotypes and proliferation of cultured cells were evaluated by morphology,immunofluorescent staining and mRNA expression of CD34,Lumican,VSX1,α-SMA and proliferating cell nuclear antigen(PCNA).in the BPE group,cells underwent serial subcultivation,and their phenotypes were identified by immunofluorescent *** analyze the proliferation of keratocytes in different concentrations of BPE,six different concentrations were designed to ascertain the most appropriate ***:In the BPE group,the cells spread out and presented dendritic morphology,and their dendrites connected to one another to form *** the third passage,most cells maintained their *** the FBS group,the cells exhibited a dendritic appearance in early cultured stages,but their morphology subsequently changed into a fibroblast-like *** number ofdendritic cells in BPE group was more than FBS and control *** staining and realtime polymerase chain reaction(PCR)confirmed that few keratocytes underwent fibroblastic transformation in the BPE and control groups,and that proliferation was higher in the BPE group than in the control *** the proliferation was higher in the FBS group,many keratocytes underwent fibroblastic *** analysis of cell morphology and mRNA expressions of CD34,PCNA and VSX1 in six group showed that different concentrations of BPE affected the proliferation obviously but didn’t affect the keratocyte phenotype,and the concentration of 40μg/mL was the most appropriate ***:BPE can improve the proliferation of keratocytes and m

关键词： keratocyte bovine pituitary extract proliferation phenotype fibroblastic transformation

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