Protocol for culturing and functionally manipulating planarian neoblasts using SiR-DNA-based flow cytometry
Protocol for culturing and functionally manipulating planarian neoblasts using SiR-DNA-based flow cytometry
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DOI：
10.1016/j.xpro.2024.102877
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发表时间：
2024-02-06
期刊：
影响因子：
--
通讯作者：
Lei，Kai
中科院分区：
文献类型：
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作者：
Zhang，Wenya;Li，Xinran;Lei，Kai
新母细胞是唯一能够在真涡虫中增殖的细胞。使用Hoechst的传统流式细胞术方案抑制细胞周期。在这里，我们提出了一个使用SiR-DNA为基础的流式细胞术的培养和功能操纵真涡虫neoblasts的协议。我们描述了细胞解离和染色、流式细胞术以及细胞收集和培养的步骤。然后，我们详细介绍了纳米荧光素酶基因转染的程序。该方案有助于进一步研究新成纤维细胞内的多能性和再生机制。关于该方案的使用和执行的完整细节，请参见Lei et al.1
Neoblasts are the only cells capable of proliferation in planarians. The traditional flow cytometry protocol using Hoechst inhibits the cell cycle. Here, we present a protocol for culturing and functionally manipulating planarian neoblasts using SiR-DNA-based flow cytometry. We describe steps for cell dissociation and staining, flow cytometry, and cell collection and culture. We then detail procedures forNanoluciferasemRNA transfection. This protocol facilitates further investigations into the pluripotency and regeneration mechanisms within neoblasts.For complete details on the use and execution of this protocol, please refer to Lei et al.1