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通过密度梯度离心,从人类肝细胞母细胞瘤组织中分离和分离各种单细胞的协议

Xianchao Zhou¹, Mei Meng¹, Yifan Wu²

¹State Key Laboratory of Systems Medicine for Cancer, Center for Single-Cell Omics, School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.

|July 17, 2023

概括

这项研究提供了一个详细的协议,用于从肝细胞瘤组织中分离高活性的肝细胞和免疫细胞,用于单细胞RNA测序. 该方法确保了用于分析肝癌异质性的高质量细胞悬浮.

科学领域:

在瘤学瘤学.
细胞生物学细胞生物学
基因组学就是基因组学.

背景情况:

单细胞转录组测序对于理解肝细胞母细胞瘤等疾病中的细胞异质性至关重要.
以前用于从肝细胞母细胞瘤组织中分离细胞的方法往往导致活力低,阻碍了下游分析.

研究的目的:

从人肝母细胞瘤样本中分离可活性肝细胞和免疫细胞的强有力的协议.
为了能够对肝细胞瘤和其他人类肝脏组织进行详细的单细胞RNA测序分析.

主要方法:

组织加工和肝细胞瘤样本的酶性消化.
珀科尔密度梯度分离以隔离特定的细胞群.
细胞悬浮质量控制和单细胞RNA (scRNA) 库的构建.

主要成果:

该协议从肝细胞瘤组织中产生高活性的单细胞悬浮体.
提供了组织解离,细胞分离和库准备的详细步骤.
该方法已通过验证,可应用于各种人类肝脏组织样本.

结论:

该协议有助于对肝细胞母细胞瘤进行全面的单细胞分析.
它提供了一种可靠的方法,用于从人类肝脏组织中制备单细胞悬浮液,用于转录基因研究.
改进的细胞隔离技术对于推进癌症研究至关重要.

关键词:

癌症癌症癌症癌症细胞隔离细胞隔离在RNAseqqq中使用.序列化是指测序的使用.一个单细胞的单细胞.

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