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

作     者：Zhong-Zhong Xu Zhi-Jie Li Lian-Xin Du Jing Li Li-Ya Wang 

作者机构：Department of Ophthalmologythe First Affiliated Hospital of Zhengzhou University Key Laboratory of Corneal DiseaseHenan Eye Institute Department of PediatricsBaylor College of Medicine 

出 版 物：《International Journal of Ophthalmology(English edition)》 (国际眼科杂志（英文版）)

年 卷 期：2013年第6卷第6期

页      面：758-765页

核心收录：

学科分类：1002[医学-临床医学] 100212[医学-眼科学] 10[医学] 

基　　金：National Natural Science Foundation of China(No.81170831) 

主　　题：keratocyte bovine pituitary extract proliferation phenotype fibroblastic transformation 

摘      要：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