Planarians possess naturally occurring pluripotent adult somatic stem cells (neoblasts) required for homeostasis and whole-body regeneration. However, no reliable neoblast culture methods are currently available, hindering mechanistic studies of pluripotency and the development of transgenic tools. We report the first robust methods for neoblast culture and delivery of exogenous mRNAs. We identify optimal culture media for maintaining neoblasts in vitro, and show via transplantation that cultured stem cells retain pluripotency for two days. We developed a new procedure that significantly improves neoblast yield and purity by modifying standard flow cytometry methods. These methods enable the introduction and expression of exogenous mRNAs in neoblasts, overcoming a key hurdle impeding the application of transgenics in planarians. The advances in cell culture reported here create new opportunities for mechanistic studies of planarian adult stem cell pluripotency, and provide a systematic framework to develop cell culture techniques in other emerging research organisms.

中文翻译：

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在涡虫干细胞培养中保留多能性和外源 mRNA 引入

涡虫拥有体内平衡和全身再生所需的天然多能成体干细胞（新生细胞）。然而，目前没有可靠的新生细胞培养方法，阻碍了多能性的机制研究和转基因工具的开发。我们报告了第一个用于新生细胞培养和外源性 mRNA 传递的稳健方法。我们确定了在体外维持新生细胞的最佳培养基，并通过移植显示培养的干细胞在两天内保持多能性。我们开发了一种新程序，通过修改标准流式细胞术方法，显着提高了新生细胞的产量和纯度。这些方法能够在新生细胞中引入和表达外源 mRNA，克服了阻碍转基因在涡虫中应用的关键障碍。本文报道的细胞培养进展为涡虫成体干细胞多能性的机制研究创造了新的机会，并为在其他新兴研究生物中开发细胞培养技术提供了系统框架。