视网膜基因移植后非人灵长类眼的制备及组织学评价。

Q1 Immunology and Microbiology

Human Gene Therapy Methods Pub Date : 2018-06-01 DOI:10.1089/hgtb.2018.045

Peter Bell, Hongwei Yu, Leah Kuntz, Omua Ahonkhai, Anna Tretiakova, Maria P Limberis, James M Wilson

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引用次数: 0

摘要

为了评估视网膜疾病的基因治疗，需要适当的人眼模型。非人类灵长类动物的眼睛比啮齿动物的眼睛有明显的优势。然而，目前的制备方法存在局限性。本文描述了基因转移后非人灵长类动物眼睛的组织学处理。使用者解剖未固定的眼睛，用滤纸将球体部分压平，并进行福尔马林固定和石蜡包埋。这种方法不需要苛刻的固定剂，允许随后的免疫染色或原位杂交，同时保留组织完整性进行组织病理学评估。此外，视网膜细胞层的垂直方向是理想的图像分析。

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Preparation of Nonhuman Primate Eyes for Histological Evaluation After Retinal Gene Transfer.

To evaluate gene therapy for retinal disorders, appropriate models of the human eye are needed. Nonhuman primate eyes offer significant advantages over rodent eyes. However, current preparation methods have limitations. Here, a protocol is described for histological processing of nonhuman primate eyes after gene transfer. The user dissects unfixed eyes, flattens the globe parts within filter paper, and performs formalin fixation and paraffin embedding. This method obviates the need for harsh fixatives, allowing subsequent immunostaining or in situ hybridization while preserving tissue integrity for histopathological evaluation. Moreover, the straight orientation of the retinal cell layers is ideal for image analysis.

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来源期刊

Human Gene Therapy Methods BIOTECHNOLOGY & APPLIED MICROBIOLOGY-GENETICS & HEREDITY

CiteScore

5.80

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Human Gene Therapy is the premier, multidisciplinary journal covering all aspects of gene therapy. The Journal publishes in-depth coverage of DNA, RNA, and cell therapies by delivering the latest breakthroughs in research and technologies. Human Gene Therapy provides a central forum for scientific and clinical information, including ethical, legal, regulatory, social, and commercial issues, which enables the advancement and progress of therapeutic procedures leading to improved patient outcomes, and ultimately, to curing diseases. The Journal is divided into three parts. Human Gene Therapy, the flagship, is published 12 times per year. HGT Methods, a bimonthly journal, focuses on the applications of gene therapy to product testing and development. HGT Clinical Development, a quarterly journal, serves as a venue for publishing data relevant to the regulatory review and commercial development of cell and gene therapy products.