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利用小鼠模型在体内诱导细胞标记并追踪标记造血细胞的方案。

Protocol to induce cell labeling in vivo and to trace labeled hematopoietic cells using mouse models.

作者信息

Syed Alina, Kobayashi Michihiro, Yoshimoto Momoko

机构信息

Center for Stem Cell and Regenerative Medicine, Brown Institute of Molecular Medicine, McGovern Medical School, University of Texas Health Science Center at Houston, Houston, TX 77030, USA.

Center for Immunobiology, Department of Investigative Medicine, Western Michigan University Homer Stryker MD School of Medicine, Kalamazoo, MI 49007, USA.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103408. doi: 10.1016/j.xpro.2024.103408. Epub 2024 Oct 21.

DOI:10.1016/j.xpro.2024.103408
PMID:39436772
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11532922/
Abstract

A combination of hematopoietic stem cell (HSC)- and endothelial cell (EC)-lineage-tracing mouse models enables us to determine blood cell origins. We present a protocol to induce cell labeling in vivo and to trace labeled hematopoietic cells to segregate their origins. We describe the steps for harvesting various hematopoietic tissues, antibody staining, and analyzing the Tomato+ percentages within each immune cell population. We also show how to estimate HSC- and EC-derived percentages of the target cell populations. For complete details on the use and execution of this protocol, please refer to Kobayashi et al..

摘要

造血干细胞（HSC）和内皮细胞（EC）谱系追踪小鼠模型的组合使我们能够确定血细胞的起源。我们提出了一种在体内诱导细胞标记并追踪标记的造血细胞以区分其起源的方案。我们描述了收获各种造血组织、抗体染色以及分析每个免疫细胞群体中番茄红素阳性（Tomato+）百分比的步骤。我们还展示了如何估计目标细胞群体中源自HSC和EC的百分比。有关此方案的使用和执行的完整详细信息，请参考小林等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eea/11532922/19301657371e/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eea/11532922/16c547fe2c2f/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eea/11532922/808aedad13d0/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eea/11532922/3c20af7293cb/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eea/11532922/19301657371e/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eea/11532922/16c547fe2c2f/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eea/11532922/808aedad13d0/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eea/11532922/3c20af7293cb/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4eea/11532922/19301657371e/gr3.jpg

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STAR Protoc. 2024 Dec 20;5(4):103408. doi: 10.1016/j.xpro.2024.103408. Epub 2024 Oct 21.
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本文引用的文献

1
HSC-independent definitive hematopoiesis persists into adult life.造血干细胞非依赖性定型造血持续存在于成年期。
Cell Rep. 2023 Mar 28;42(3):112239. doi: 10.1016/j.celrep.2023.112239. Epub 2023 Mar 11.
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Murine HSCs contribute actively to native hematopoiesis but with reduced differentiation capacity upon aging.鼠类造血干细胞在体内造血中发挥积极作用，但随着年龄的增长其分化能力会下降。
Elife. 2018 Dec 18;7:e41258. doi: 10.7554/eLife.41258.
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Fgd5 identifies hematopoietic stem cells in the murine bone marrow.Fgd5可识别小鼠骨髓中的造血干细胞。
J Exp Med. 2014 Jun 30;211(7):1315-31. doi: 10.1084/jem.20130428. Epub 2014 Jun 23.
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Ephrin-B2 controls VEGF-induced angiogenesis and lymphangiogenesis.Ephrin-B2 控制 VEGF 诱导的血管生成和淋巴管生成。
Nature. 2010 May 27;465(7297):483-6. doi: 10.1038/nature09002.