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学科主题: 临床医学
题名:
Glial cell-derived neurotrophic factor gene delivery enhances survival of human corneal epithelium in culture and the overexpression of GDNF in bioengineered constructs
作者: Qi, Hong1,4; Shine, H. David2,3; Li, De-Quan1; de Paiva, Cintia S.1; Farley, William J.1; Jones, Dan B.1; Pflugfelder, Stephen C.1
关键词: cornea ; epithelium ; glial cell-derived neurotrophic factor ; gene therapy ; adenoviral vector ; bioengineering
刊名: EXPERIMENTAL EYE RESEARCH
发表日期: 2008-12-01
DOI: 10.1016/j.exer.2008.09.012
卷: 87, 期:6, 页:580-586
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Ophthalmology
研究领域[WOS]: Ophthalmology
关键词[WOS]: EXPANDED EX-VIVO ; TARGETED TRANSDUCTION ; TRANSGENE EXPRESSION ; HUMAN LIMBAL ; STEM-CELLS ; IN-VIVO ; SURFACE ; LINE ; TRANSPLANTATION ; THERAPY
英文摘要:

This paper evaluates the effects of adenoviral vector-mediated glial cell-derived neurotrophic factor (GDNF) gene delivery on survival of primary human corneal epithelial cells (PHCEC) established from limbal explants in vitro and the overexpression of GDNF gene in bioengineered human corneal constructs on substrate of corneal stromal discs followed by autograft ex vivo. In vitro, the overexpression of GDNF in the supernatant of PHCEC peaked at day 4, but lasted for at least 4 weeks after the transduction mediated by adenoviral vector. At day 10, the cell viability was 2-fold greater (P < 0.001), the number of terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL)-positive cells was more than 50% lower (P < 0.01) in the GDNF transduction group than the non-transduction group. 5 weeks after the transduction, the living cell population was greater in the GDNF transduction group than the non-transduction group (P < 0.01). In the ex vivo autograft of the bioengineered human corneal constructs, outgrowth of enhanced green fluorescent protein (eGFP) positive cells on the recipient corneoscleral tissue was observed. Overexpression of GDNF in the supernatant peaked at day 2, but was observed for at least 4 weeks after transplantation. At day 5, immunofluorescent staining showed expression of GDNF by all layers of epithelial cells on the graft. Our findings revealed that GDNF is a survival growth factor for cultured human corneal epithelium. The use of bioengineered human corneal constructs containing GDNF-transduced epithelial cells represents a novel method for delivering of this gene to promote survival of transplanted corneal epithelium to treat various corneal surface diseases. (C) 2008 Elsevier Ltd. All rights reserved.

语种: 英语
所属项目编号: EY11915 ; NS35280 ; EY014553 ; PRMRP FY06 PR064719
项目资助者: NIH ; National Institutes of Health, Bethesda, MD ; Department of Defense Congressionally Directed Medical Research Programs (CDMRP) ; Lions Eye Bank Foundation ; Research to Prevent Blindness ; Oshman Foundation ; William Stamps Farish Fund
WOS记录号: WOS:000261823900012
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/56378
Appears in Collections:北京大学第三临床医学院_眼科_期刊论文

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作者单位: 1.Baylor Coll Med, Dept Ophthalmol, Ocular Surface Ctr, Cullen Eye Inst, Houston, TX 77030 USA
2.Baylor Coll Med, Ctr Cell & Gene Therapy, Houston, TX 77030 USA
3.Baylor Coll Med, Dept Neurosurg, Houston, TX 77030 USA
4.Peking Univ, Peking Univ Hosp 3, Peking Univ Eye Ctr, Beijing 100871, Peoples R China

Recommended Citation:
Qi, Hong,Shine, H. David,Li, De-Quan,et al. Glial cell-derived neurotrophic factor gene delivery enhances survival of human corneal epithelium in culture and the overexpression of GDNF in bioengineered constructs[J]. EXPERIMENTAL EYE RESEARCH,2008,87(6):580-586.
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