纯化小鼠肝脏非实质细胞或细胞核，用于染色质免疫沉淀测序（ChIP-seq）及其他新一代测序方法。

Troutman Ty D, Bennett Hunter, Sakai Mashito, Seidman Jason S, Heinz Sven, Glass Christopher K

Department of Medicine, University of California, San Diego, La Jolla, CA 92093, USA.

Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA.

STAR Protoc. 2021 Mar 10;2(1):100363. doi: 10.1016/j.xpro.2021.100363. eCollection 2021 Mar 19.

Significant advancements in understanding disease mechanisms can occur through combined analysis of next-generation sequencing datasets generated using purified cell populations. Here, we detail our optimized protocol for purification of mouse hepatic macrophages (or other liver non-parenchymal populations) suitable for use in various next-generation sequencing protocols. An alternative framework is described for sorting pre-fixed hepatic nuclei populations. This strategy has the advantage of rapidly preserving the nuclei and can facilitate success with ChIP-seq for more challenging molecules. For complete details on the use and execution of these protocols, please refer to Muse et al. (2018), Sakai et al. (2019), and Seidman et al. (2020).

通过对使用纯化细胞群体生成的下一代测序数据集进行联合分析，在理解疾病机制方面可以取得重大进展。在这里，我们详细介绍了我们优化的方案，用于纯化适用于各种下一代测序方案的小鼠肝巨噬细胞（或其他肝非实质细胞群体）。还描述了一种用于分选预固定肝细胞核群体的替代框架。这种策略具有快速保存细胞核的优点，并且可以促进对更具挑战性分子进行ChIP-seq的成功。有关这些方案的使用和执行的完整详细信息，请参考Muse等人（2018年）、Sakai等人（2019年）和Seidman等人（2020年）的文献。