使用牛的垂体摘录增加 keratocytes 的增长并且在 vitro 维持他们的显型

• 作者: Jing Li Li-Ya Wang Lian-Xin Du Zhi-Jie Li Zhong-Zhong Xu
• 来源期刊: 国际眼科杂志：英文版
• 学科: 医学
• 关键词: KERATOCYTE BOVINE PITUITARY extract proliferation PHENOTYPE fibroblastic transformation
• 年，卷（期）: 2013,（6）
• 页码范围: 758-765
• 页数: 8页
• 分类号: R772

摘要

AIM:To investigate the effects of bovine pituitary extract on the proliferation of keratocytes and maintaining the keratocyte phenotype/n v/tro.METHODS:Single keratocytes were isolated by enzyme digestion for in vitro culture.Three groups were designed according to the different culture media:a bovine pituitary extract(BPE)group,a fetal bovine serum(FBS)group and the control group.The 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 staining.To analyze the proliferation of keratocytes in different concentrations of BPE,six different concentrations were designed to ascertain the most appropriate amount.RESULTS:In the BPE group,the cells spread out and presented dendritic morphology,and their dendrites connected to one another to form networks.On the third passage,most cells maintained their phenotype.In the FBS group,the cells exhibited a dendritic appearance in early cultured stages,but their morphology subsequently changed into a fibroblast-like shape.The number ofdendritic cells in BPE group was more than FBS and control groups.Immunofluorescent 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 group.Although the proliferation was higher in the FBS group,many keratocytes underwent fibroblastic transformation.The 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 one.CONCLUSION:BPE can improve the proliferation of keratocytes and maintain their phenotype in vitro.Many kerat