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相关实验视频

Updated: Jul 25, 2025

Isolation of Nuclei from Flash-Frozen Liver Tissue for Single-Cell Multiomics
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[对来自小鼠肝脏的多种类型细胞的同步分离方法]

J Gan1, C F Ji1, X R Mao1

  • 1The First Clinical Medical College of Lanzhou University, Lanzhou 730000, China.

Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
|June 26, 2023
PubMed
概括

这项研究提出了一种有效的方法,用于从小鼠肝脏中分离肝细胞,肝星细胞 (HSC) 和淋巴细胞. 该技术确保了高细胞活力和纯度,简化了肝细胞研究.

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科学领域:

  • 肝病学和免疫学 肝病学和免疫学
  • 细胞生物学 细胞生物学
  • 生物技术是生物技术.

背景情况:

  • 隔离多个主要肝细胞,包括肝细胞,肝星细胞 (HSC) 和淋巴细胞,对于研究肝功能和疾病至关重要.
  • 现有的方法可能很复杂,耗时,需要更简单,更高效的技术.

研究的目的:

  • 开发和验证一种简单有效的方法,同时隔离和净化来自小鼠肝脏的肝细胞,肝细胞和淋巴细胞.
  • 评估分离细胞的活力和纯度.

主要方法:

  • 使用肝输液与门静脉消化,从C57bl/6小鼠中获得细胞悬浮液.
  • 采用不连续的Percoll梯度离心法进行细胞隔离和净化.
  • 评估细胞活力使用试剂蓝色排除;通过糖原染色,细胞克拉18和电子显微镜识别肝细胞;通过α-光滑肌动因和脱因的免疫光学来表征HCs;通过流细胞计分析淋巴细胞子集.

主要成果:

  • 从小鼠肝脏中成功分离出大约2.7×10^7个肝细胞,5.7×10^5个肝细胞和4.6×10^6个肝单核细胞.
  • 在所有孤立的细胞类型中,细胞存活率超过95%.
  • 通过特定的标记物和技术,通过特定的标记物和技术确认了肝细胞,肝细胞细胞和各种淋巴细胞子集 (CD4,CD8,NK,NKT) 的身份和纯度.
关键词:
肝脏的星状细胞是什么肝细胞 肝细胞 肝细胞隔离 隔离 隔离 隔离淋巴细胞 淋巴细胞初级文化是一种初级文化.

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结论:

  • 描述的肝输液和门静脉消化方法提供了一种简单,高效和同时的方法,用于从小鼠中分离多个原发性肝细胞.
  • 这种技术对于需要多样化的肝细胞群体的研究是有价值的.