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Lgr5肠道干细胞分选及类器官培养。

Lgr5 intestinal stem cell sorting and organoid culture.

作者信息

Liu Ruixue, Li Haifeng, Cai Juan, Wei Qiang, Han Xiaonan

机构信息

Key Laboratory of Human Disease Comparative Medicine The Ministry of Health Institute of Laboratory Animal Science (ILAS) Chinese Academy of Medical Science (CAMS) and Peking Union Medical College (PUMC) Beijing P.R. China.

出版信息

Animal Model Exp Med. 2019 Mar 12;2(2):132-135. doi: 10.1002/ame2.12057. eCollection 2019 Jun.

DOI:10.1002/ame2.12057
PMID:31392307
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6600610/
Abstract

Intestinal epithelial stem cells (IESCs) are one of the most rapidly self-renewing and proliferating adult stem cells. The IESCs reside at the bottom of intestinal and colonic crypts, giving rise to all intestinal epithelial lineages and maintaining intestinal epithelial replenishment. The technique of three-dimensional culture based upon intestinal stem cell biology has been recently developed to study gastrointestinal development and disease pathogenesis. Here, we summarize the techniques used to isolate Lgr5-positive IESCs to form the enteroids from intestine or colonoids from colon, and present the means to examine these organoid functions. This study will provide a simple and practical way for producing intestinal tissues in the laboratory.

摘要

肠道上皮干细胞(IESCs)是自我更新和增殖速度最快的成体干细胞之一。IESCs 位于小肠和结肠隐窝底部,可分化为所有肠道上皮谱系并维持肠道上皮细胞的更新。基于肠道干细胞生物学的三维培养技术最近已被开发出来,用于研究胃肠道发育和疾病发病机制。在这里,我们总结了用于从小肠中分离 Lgr5 阳性 IESCs 以形成肠类器官或从结肠中分离 Lgr5 阳性 IESCs 以形成结肠类器官的技术,并介绍了检测这些类器官功能的方法。本研究将为在实验室中生产肠道组织提供一种简单实用的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8cb8/6600610/f3d05186b5dc/AME2-2-132-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8cb8/6600610/51a4845b0712/AME2-2-132-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8cb8/6600610/562e39336db2/AME2-2-132-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8cb8/6600610/f3d05186b5dc/AME2-2-132-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8cb8/6600610/51a4845b0712/AME2-2-132-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8cb8/6600610/562e39336db2/AME2-2-132-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8cb8/6600610/f3d05186b5dc/AME2-2-132-g003.jpg

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本文引用的文献

1
Intestinal Enteroendocrine Lineage Cells Possess Homeostatic and Injury-Inducible Stem Cell Activity.肠道肠内分泌谱系细胞具有稳态和损伤诱导的干细胞活性。
Cell Stem Cell. 2017 Jul 6;21(1):78-90.e6. doi: 10.1016/j.stem.2017.06.014.
2
Induced Quiescence of Lgr5+ Stem Cells in Intestinal Organoids Enables Differentiation of Hormone-Producing Enteroendocrine Cells.诱导肠类器官中 Lgr5+干细胞静息使其能够分化为产生激素的肠内分泌细胞。
Cell Stem Cell. 2017 Feb 2;20(2):177-190.e4. doi: 10.1016/j.stem.2016.11.001. Epub 2016 Dec 8.
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Activated STAT5 confers resistance to intestinal injury by increasing intestinal stem cell proliferation and regeneration.
使用光激活细胞染料进行空间转录组学的活细胞标记、追踪和分离。
Nat Commun. 2021 Aug 17;12(1):4995. doi: 10.1038/s41467-021-25279-y.
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Microbiol Spectr. 2021 Sep 3;9(1):e0000321. doi: 10.1128/Spectrum.00003-21. Epub 2021 Jun 9.
激活的 STAT5 通过增加肠道干细胞的增殖和再生赋予肠道免受损伤的能力。
Stem Cell Reports. 2015 Feb 10;4(2):209-25. doi: 10.1016/j.stemcr.2014.12.004. Epub 2015 Jan 8.
4
Mapping early fate determination in Lgr5+ crypt stem cells using a novel Ki67-RFP allele.利用新型 Ki67-RFP 等位基因绘制 Lgr5+ 隐窝干细胞的早期命运决定。
EMBO J. 2014 Sep 17;33(18):2057-68. doi: 10.15252/embj.201488017. Epub 2014 Aug 4.
5
The pan-ErbB negative regulator Lrig1 is an intestinal stem cell marker that functions as a tumor suppressor.pan-ErbB 阴性调节剂 Lrig1 是一种肠道干细胞标志物,具有肿瘤抑制作用。
Cell. 2012 Mar 30;149(1):146-58. doi: 10.1016/j.cell.2012.02.042.
6
Interconversion between intestinal stem cell populations in distinct niches.不同龛位肠道干细胞群体之间的相互转换。
Science. 2011 Dec 9;334(6061):1420-4. doi: 10.1126/science.1213214. Epub 2011 Nov 10.
7
A reserve stem cell population in small intestine renders Lgr5-positive cells dispensable.小肠中的储备干细胞群体使 Lgr5 阳性细胞变得可有可无。
Nature. 2011 Sep 18;478(7368):255-9. doi: 10.1038/nature10408.
8
Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells.肠隐窝稳态源自对称分裂的 Lgr5 干细胞之间的中性竞争。
Cell. 2010 Oct 1;143(1):134-44. doi: 10.1016/j.cell.2010.09.016.
9
Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche.单个Lgr5干细胞在体外无需间充质微环境即可构建隐窝-绒毛结构。
Nature. 2009 May 14;459(7244):262-5. doi: 10.1038/nature07935. Epub 2009 Mar 29.
10
Bmi1 is expressed in vivo in intestinal stem cells.Bmi1在肠道干细胞中在体内表达。
Nat Genet. 2008 Jul;40(7):915-20. doi: 10.1038/ng.165. Epub 2008 Jun 8.