Dissociation and flow cytometric isolation of murine intestinal epithelial cells for multi-omic profiling.
Dissociation and flow cytometric isolation of murine intestinal epithelial cells for multi-omic profiling.
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DOI：
10.1016/j.xpro.2022.101936
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发表时间：
2023-03-17
期刊：
影响因子：
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通讯作者：
Mohamadzadeh, Mansour
中科院分区：
文献类型：
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作者：
Ge, Yong;Zadeh, Mojgan;Mohamadzadeh, Mansour
肠上皮由多种细胞类型组成，这些细胞可以分离，但由于失巢凋亡而难以维持高细胞活力。在此，我们描述了使用乙二胺四乙酸和TrypLE Express分离高活力肠上皮细胞的分步方案，随后可用于多组学分析，包括单细胞RNA测序。有关本方案使用和执行的完整详细信息，请参见Ge等人。（2022年）。我们描述了通过流式细胞术对EC进行染色和分析的程序。纯化的EC可用于多组学分析，包括scRNA-seq。肠上皮细胞是由多种细胞组成的，这些细胞可以分离，但由于失巢凋亡而难以保持高的细胞活力。在此，我们描述了一个逐步的协议，用于分离高活力的肠上皮细胞，使用乙二胺四乙酸和TrypLE表达，随后可用于多组学分析，包括单细胞RNA测序。
Intestinal epithelium is composed of several cell types, which can be dissociated but difficult to maintain high cell viability due to anoikis. Herein, we describe a step-by-step protocol for the isolation of highly viable intestinal epithelial cells using ethylenediaminetetraacetate acid and TrypLE Express, which can subsequently be employed for multi-omic analyses, including single-cell RNA sequencing. For complete details on the use and execution of this protocol, please refer to Ge et al. (2022). Dissociation of murine ECs from intestine We describe the procedures to stain and analyze ECs by flow cytometry Purified ECs can be used for multi-omic analysis, including scRNA-seq Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Intestinal epithelium is composed of several cell types, which can be dissociated but difficult to maintain high cell viability due to anoikis. Herein, we describe a step-by-step protocol for the isolation of highly viable intestinal epithelial cells using ethylenediaminetetraacetate acid and TrypLE Express, which can subsequently be employed for multi-omic analyses, including single-cell RNA sequencing.
影响因子：
5.8
作者：
Ge, Yong;Zadeh, Mojgan;Yang, Changjun;Candelario-Jalil, Eduardo;Mohamadzadeh, Mansour
通讯作者：
Mohamadzadeh, Mansour
DOI：
10.1084/jem.20220057
发表时间：
2022-07-04
期刊：
The Journal of experimental medicine
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