诱导多能干细胞来源的视网膜色素上皮培养物中drusen的免疫荧光表征方法。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-06-20 Epub Date: 2025-05-02 DOI:10.1016/j.xpro.2025.103791

Sonal Dalvi, Ruchira Singh

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引用次数: 0

摘要

Drusen是视网膜色素上皮（RPE）下富含脂蛋白的沉积物，是老年性黄斑变性和相关黄斑营养不良的病理标志。在这里，我们描述了一种在诱导多能干细胞(iPSC)-RPE （iRPE）单培养下观察积水沉积物的方案。我们描述了iRPE细胞培养的步骤，随后，酶去除iRPE单层和亚rpe沉淀的免疫染色。然后，我们详细步骤的共聚焦显微镜成像免疫染色的dren在iRPE培养。有关本协议使用和执行的完整细节，请参阅Dalvi等人1。

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Protocol for immunofluorescence characterization of drusen in induced pluripotent stem cell-derived retinal pigment epithelium cultures.

Drusen, lipoprotein-rich deposits underlying retinal pigment epithelium (RPE), are a pathological hallmark of age-related macular degeneration and related macular dystrophies. Here, we describe a protocol for visualizing drusen deposits beneath induced pluripotent stem cell (iPSC)-RPE (iRPE) monocultures. We describe steps for iRPE cell culture and, subsequently, enzymatic removal of the iRPE monolayer and immunostaining of sub-RPE drusen deposits. We then detail steps for confocal microscopy imaging of immunostained drusen in iRPE culture. For complete details on the use and execution of this protocol, please refer to Dalvi et al.¹.

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