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使用低剂量阿糖胞苷将原代视网膜祖细胞分化为视网膜神经节样细胞。

Differentiation of primary retinal progenitor cells into retinal ganglion-like cells using low dose cytarabine.

作者信息

Hu Baoqi, Zhou Sijia, Wang Xiou, Zhang Zhichao, Wang Rui, Kang Qianyan

机构信息

Department of Ophthalmology, Xi'an No.1 Hospital, the First Affiliated Hospital of Northwest University, Xi'an, Shaanxi, 710002, China; Department of Ophthalmology, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, China; Shaanxi Institute of Ophthalmology, Xi'an, Shaanxi, 710002, China.

Department of Ophthalmology, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, China.

出版信息

Biochem Biophys Res Commun. 2025 Feb 8;748:151359. doi: 10.1016/j.bbrc.2025.151359. Epub 2025 Jan 17.

Abstract

The death of retinal ganglion cells (RGCs) is a key factor in the pathophysiology of all forms of glaucoma. RGC culture serves as a simple system for establishing and testing candidate therapies. This study aimed to explore the differentiation of primary retinal progenitor cells (RPCs) into RGC-like cells induced by low-dose cytarabine (Ara-C). RPCs were isolated from the retina of newborn rats and cultured in vitro. Different concentrations of Ara-C were added to the culture medium to induce the differentiation of RPCs into RGC-like cells. Differentiation efficiency was assessed through immunofluorescence staining and cell counting. The addition of Ara-C significantly increased the number of Brn3a/RBPMS double-positive cells. The RPC-RGCs induced displayed characteristic features of RGCs, with roughly 80.9 % ± 6.2 % of the cells positive for both TuJ1/NeuN and 77.5 % ± 4.9 % for Brn3a/RBPMS. The study demonstrates that the addition of Ara-C to primary cultures of rat RPCs can enhance their differentiation into RGC-like cells, providing a simple and rapid method for obtaining RGC-like cells with a relatively high purity. This method shows considerable promise for advancing glaucoma research and potential therapeutic strategies to restore vision after RGC loss.

摘要

视网膜神经节细胞(RGCs)的死亡是所有形式青光眼病理生理学的关键因素。RGC培养是建立和测试候选疗法的简单系统。本研究旨在探讨低剂量阿糖胞苷(Ara-C)诱导原代视网膜祖细胞(RPCs)向RGC样细胞的分化。从新生大鼠视网膜中分离RPCs并进行体外培养。向培养基中添加不同浓度的Ara-C以诱导RPCs向RGC样细胞分化。通过免疫荧光染色和细胞计数评估分化效率。添加Ara-C显著增加了Brn3a/RBPMS双阳性细胞的数量。诱导的RPC-RGCs表现出RGCs的特征,约80.9%±6.2%的细胞TuJ1/NeuN双阳性,77.5%±4.9%的细胞Brn3a/RBPMS双阳性。该研究表明,向大鼠RPCs原代培养物中添加Ara-C可增强其向RGC样细胞的分化,为获得相对高纯度的RGC样细胞提供了一种简单快速的方法。该方法在推进青光眼研究和RGC损失后恢复视力的潜在治疗策略方面显示出巨大前景。

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