Isolation of satellite cells and transplantation into mice for lineage tracing in muscle.,Nature Protocols

当前位置： X-MOL 学术 › Nat. Protoc. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Isolation of satellite cells and transplantation into mice for lineage tracing in muscle.
Nature Protocols ( IF 13.1 ) Pub Date : 2020-01-22 , DOI: 10.1038/s41596-019-0278-8
Peter Feige _{1,

2} , Michael A Rudnicki _{1,

2,

3}

Affiliation

Limited methods exist to assay the direct effects of therapeutic intervention on muscle stem cell fate, proliferation or differentiation in an in vivo context. Here we provide an optimized protocol for muscle stem cell isolation and transplantation into mice to deconvolute heterogeneity within isolated stem cell populations. Viable and pure cell populations are isolated within 2 h and can then be used for therapeutic intervention or transplantation to uncover the repopulating and differentiation potential in mice, a physiologically relevant in vivo context. Effects can be assessed 9 d after transplantation. This methodology analyzes cell and sort purity prior to transplantation to improve reproducibility and outlines novel blocking steps to improve tissue staining and analysis. Experience with surgical procedures in mice is recommended before attempting this protocol. Our system is widely applicable for exploring stem cell dynamics within muscle and has already been used to study heterogeneity within muscle stem cell populations and efficacy of therapeutic intervention on isolated stem cell populations.

中文翻译：

分离卫星细胞并将其移植到小鼠中以在肌肉中进行谱系追踪。

存在有限的方法来测定体内背景下治疗干预对肌肉干细胞命运，增殖或分化的直接作用。在这里，我们为肌肉干细胞分离和移植到小鼠中以优化卷积异质性分离的干细胞群体中提供了一个优化的协议。在2小时内分离出了存活的纯细胞群体，然后可将其用于治疗干预或移植，以发现小鼠（在生理上相关的体内情况）的再增殖和分化潜能。移植后9 d可评估疗效。该方法在移植前分析细胞和分选纯度以提高可重复性，并概述了新的阻断步骤以改善组织染色和分析。建议在尝试此方案之前，先对小鼠进行外科手术，以丰富经验。我们的系统可广泛用于探索肌肉内的干细胞动力学，并且已经用于研究肌肉干细胞群内的异质性以及对孤立的干细胞群进行治疗性干预的功效。

更新日期：2020-01-22

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11