Gordon Tal, Hendin Noam, Wurtzel Omri
The School of Neurobiology, Biochemistry and Biophysics, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel.
Sagol School of Neuroscience, Tel Aviv University, Tel Aviv, Israel.
Front Cell Dev Biol. 2023 Oct 11;11:1274826. doi: 10.3389/fcell.2023.1274826. eCollection 2023.
is the only molecularly characterized solitary ascidian capable of regenerating all organs and tissue types. The cellular basis for regeneration in is largely unknown, and methods for isolating live cells from this species for functional analyses are unavailable. Here, we developed a method for isolating live cells from , overcoming major experimental challenges, including the dissociation of its thick body wall and native cellular autofluorescence. We demonstrated the applicability of our approach for tissue dissociation and cell analysis using three flow cytometry platforms, and by using broadly used non-species-specific cell labeling reagents. In addition to live cell isolation, proof-of-concept experiments showed that this approach was compatible with gene expression analysis of RNA extracted from the isolated cells, and with analysis of phagocytosis. We presented efficient methods for cell purification from a highly regenerative ascidian, which could be transferable to diversity of non-model marine organisms. The ability to purify live cells will promote future studies of cell function in regeneration.
是唯一一种在分子层面有特征描述的能够再生所有器官和组织类型的独居海鞘。其再生的细胞基础在很大程度上尚不清楚，且目前尚无从该物种中分离活细胞用于功能分析的方法。在此，我们开发了一种从[物种名称]中分离活细胞的方法，克服了主要的实验挑战，包括其厚体壁的解离和天然细胞自发荧光。我们使用三个流式细胞术平台，并通过使用广泛使用的非物种特异性细胞标记试剂，证明了我们的方法在组织解离和细胞分析中的适用性。除了活细胞分离，概念验证实验表明该方法与从分离细胞中提取的RNA的基因表达分析以及吞噬作用分析兼容。我们展示了从一种高度可再生的海鞘中进行细胞纯化的有效方法，该方法可能可转移到多种非模式海洋生物中。纯化活细胞的能力将促进未来对[物种名称]再生中细胞功能的研究。
