Bell Peter, Yu Hongwei, Kuntz Leah, Ahonkhai Omua, Tretiakova Anna, Limberis Maria P, Wilson James M
Gene Therapy Program, Department of Medicine, Perelman School of Medicine, University of Pennsylvania , Philadelphia, Pennsylvania.
Hum Gene Ther Methods. 2018 Jun;29(3):115-123. doi: 10.1089/hgtb.2018.045.
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.
为评估视网膜疾病的基因治疗效果,需要合适的人眼模型。与啮齿动物眼睛相比,非人灵长类动物眼睛具有显著优势。然而,目前的制备方法存在局限性。在此,描述了一种基因转移后非人灵长类动物眼睛组织学处理的方案。使用者解剖未固定的眼睛,将眼球部分在滤纸上展平,然后进行福尔马林固定和石蜡包埋。该方法无需使用强固定剂,在保留组织完整性以进行组织病理学评估的同时,允许后续的免疫染色或原位杂交。此外,视网膜细胞层的直线排列非常适合图像分析。