Hu Baoqi, Zhou Sijia, Wang Xiou, Zhang Zhichao, Wang Rui, Kang Qianyan
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.
Biochem Biophys Res Commun. 2025 Feb 8;748:151359. doi: 10.1016/j.bbrc.2025.151359. Epub 2025 Jan 17.
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.
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。
