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

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

phenotype in vitro

Zhong-Zhong Xu; Zhi-Jie Li; Lian-Xin Du; Jing Li; Li-Ya Wang

【期刊名称】《《国际眼科杂志：英文版》》 【年(卷),期】2013(000)006

【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 antigen(PCNA).in subcultivation,and

the their

and BPE

proliferating group,cells

cell

nuclear serial by

underwent

identified

phenotypes were

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 keratocytes can be harvested rapidly and provide seeds for the construction of corneal stroma. 【总页数】8页(P.758-765)

【关键词】keratocyte bovine pituitary extract proliferation phenotype fibroblastic transformation

【作者】Zhong-Zhong Xu; Zhi-Jie Li; Lian-Xin Du; Jing Li; Li-Ya Wang 【作者单位】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 【正文语种】中文 【中图分类】R772 【文献来源】

https://www.zhangqiaokeyan.com/academic-journal-cn_international-journal-ophthalmology_thesis/0201273696014.html 【相关文献】

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