NASH is a chronic liver disease that affects 3%–6% of individuals and requires urgent therapeutic developments. Isolating the key cell types in the liver is a necessary step towards understanding their function and roles in disease pathogenesis. However, traditional isolation methods through gradient centrifugation can only collect one or a few cell types simultaneously and pose technical difficulties when applied to NASH livers. Taking advantage of identified cell surface markers from liver single-cell RNAseq, here we established the combination of gradient centrifugation and antibody-based cell sorting techniques to isolate five key liver cell types (hepatocytes, endothelial cells, stellate cells, macrophages and other immune cells) from a single mouse liver. This method yielded high purity of each cell type from healthy and NASH livers. Our five-in-one protocol simultaneously isolates key liver cell types with high purity under normal and NASH conditions, enabling for systematic and accurate exploratory experiments such as RNA sequencing.

中文翻译：

---

五合一：从正常和 NASH 小鼠的肝脏中同时分离多种主要肝细胞类型

NASH 是一种慢性肝病，影响 3%–6% 的个体，需要紧急治疗开发。分离肝脏中的关键细胞类型是了解它们在疾病发病机制中的功能和作用的必要步骤。然而，传统的梯度离心分离方法只能同时收集一种或几种细胞类型，在应用于 NASH 肝脏时存在技术困难。利用肝脏单细胞 RNAseq 中已鉴定的细胞表面标志物，我们建立了梯度离心和基于抗体的细胞分选技术的组合，以分离五种关键的肝细胞类型（肝细胞、内皮细胞、星状细胞、巨噬细胞和其他免疫细胞） ) 来自单个小鼠肝脏。这种方法从健康和 NASH 肝脏中产生了高纯度的每种细胞类型。