从小鼠脂肪肝中分离原代肝细胞和非实质细胞
Isolation of primary hepatocytes and non-parenchymal cells from mouse fatty liver.
摘要(中文译文)
在代谢相关脂肪性肝病（MAFLD）的早期阶段，脂质蓄积发生于肝脏的实质细胞（肝细胞，HPCs），表现为肝细胞内脂滴数量增加以及细胞密度整体下降。不同肝细胞类型之间的细胞间通讯影响肝脏脂质蓄积，使得非实质细胞（NPCs）对HPCs的影响尤为重要。体外HPC模型已成熟建立并广泛应用于MAFLD研究。然而，MAFLD中微环境的改变以及HPCs与NPCs之间的交叉污染，使得单一细胞类型中的基因表达研究不够精确。为更精确地检测两种细胞类型之间的差异基因表达，我们在采用灌注法提取HPCs时加强了清洗细节，并在提取NPCs时将Percoll浓度梯度调整为25%/60%。与先前方法相比，我们的结果显示，HFD（高脂饮食）小鼠肝脏中HPCs的细胞活力更高，达94.3%，NPCs为91.4%。此外，HPCs和NPCs的纯度也更高。这一方法学改进使得能够准确定量细胞间基因表达差异，为阐明细胞间串扰在MAFLD发病机制中的作用开辟了新途径。
Abstract
In the early stages of metabolism-associated fatty liver disease (MAFLD), lipid accumulation occurs in the parenchymal cells (hepatocytes, HPCs) of the liver, manifested by an increase in the number of lipid droplets within hepatocytes and an overall decrease in cellular density. Intercellular communication among different liver cell types influences hepatic lipid accumulation, making the impact of non-parenchymal cells (NPCs)on HPCs particularly significant. In vitro HPC models have been well-established and widely applied in MAFLD research. However, the altered microenvironment in MAFLD and cross-contamination between HPCs and NPCs make gene expression research in a single cell type imprecise. To more precisely detect differential gene expression between the two cell types, we enhanced cleaning details during HPC extraction using the perfusion method and adjusted the Percoll concentration gradient to 25%/60% during NPCs extraction. Compared with the previous method, our results show higher cell viability of 94.3% for HPCs and 91.4% for NPCs in the livers of HFD (High Fat Diet) mice. Furthermore, the purity of HPCs and NPCs is higher. This methodological refinement enables accurate quantification of intercellular differences in gene expression, opening new avenues to delineate the role of cell-cell crosstalk in MAFLD pathogenesis.
如何引用
AMA
Xinzhe Zhuo, Kai Wang, Yunan Shi, Xiang Wang, Huabing Zhan, Jiao Yu, et al. Isolation of primary hepatocytes and non-parenchymal cells from mouse fatty liver.. MethodsX. 2026; doi:10.1016/j.mex.2026.103888.
APA
Xinzhe Zhuo, Kai Wang, Yunan Shi, Xiang Wang, Huabing Zhan, Jiao Yu, et al (2026). Isolation of primary hepatocytes and non-parenchymal cells from mouse fatty liver.. MethodsX. https://doi.org/10.1016/j.mex.2026.103888
GB/T 7714
Xinzhe Zhuo, Kai Wang, Yunan Shi, Xiang Wang, Huabing Zhan, Jiao Yu, et al. Isolation of primary hepatocytes and non-parenchymal cells from mouse fatty liver.[J]. MethodsX, 2026 doi:10.1016/j.mex.2026.103888.
在 PubMed增强版 阅读全文
翻译全文 · AI 文献问答 · PDF 原文下载 · 引用导出