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人诱导多能干细胞中单核细胞和极化 M1/M2 巨噬细胞的分化。

Differentiation of monocytes and polarized M1/M2 macrophages from human induced pluripotent stem cells.

机构信息

Ocular Stem Cells and Translational Research (OSCTR) Section, Ophthalmic Genetic and Visual Function Branch (OGVFB), National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.

Ocular Stem Cells and Translational Research (OSCTR) Section, Ophthalmic Genetic and Visual Function Branch (OGVFB), National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA.

出版信息

STAR Protoc. 2024 Mar 15;5(1):102827. doi: 10.1016/j.xpro.2023.102827. Epub 2024 Jan 13.

DOI:10.1016/j.xpro.2023.102827
PMID:38219151
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10826317/
Abstract

Here, we present a protocol to differentiate induced pluripotent stem cell (iPSC) into adherent hematopoietic progenitors that release floating CD14+ CD45+ monocytes into the culture medium. We describe steps for iPSC expansion, embryoid body (EB) formation, suspension culture, plating EBs, and recurring harvests of monocytes, a.k.a. "monocyte factory." We then describe detailed procedures for freezing/thawing of monocytes and differentiation into polarized M1 and M2 macrophages. This protocol provides foundation to study iPSC monocytes and their progenies such as macrophages, microglial, and dendritic cells. For complete details on the use and execution of this protocol, please refer to Karlson et al. and Panicker et al..

摘要

在此，我们提供了一个方案，可将诱导多能干细胞（iPSC）分化为贴壁造血祖细胞，这些祖细胞将漂浮的 CD14+ CD45+ 单核细胞释放到培养基中。我们描述了 iPSC 扩增、类胚体（EB）形成、悬浮培养、EB 接种以及单核细胞（也称为“单核细胞工厂”）的反复收获的步骤。然后，我们描述了详细的单核细胞冻融和分化为极化 M1 和 M2 巨噬细胞的程序。该方案为研究 iPSC 单核细胞及其祖细胞（如巨噬细胞、小神经胶质细胞和树突状细胞）提供了基础。有关此方案的使用和执行的完整详细信息，请参考 Karlson 等人和 Panicker 等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/b8764a4e2307/gr10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/0756dff81c30/fx1.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/1329af5a8df1/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/4f6240091112/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/b1added1d228/gr4.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/1864f4edf84c/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/837c1b5f2b6e/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/26d8cf81232c/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/b8764a4e2307/gr10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/0756dff81c30/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/cf0d701e892d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/1329af5a8df1/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/4f6240091112/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/b1added1d228/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/46fe658587ae/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/c27eac7be2d1/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/1864f4edf84c/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/837c1b5f2b6e/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/26d8cf81232c/gr9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bf5/10826317/b8764a4e2307/gr10.jpg

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Differentiation of human induced pluripotent stem cells to authentic macrophages using a defined, serum-free, open-source medium.使用一种定义明确、无血清、开源的培养基将人诱导多能干细胞分化为真正的巨噬细胞。
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High-Yield Human Induced Pluripotent Stem Cell-Derived Monocytes and Macrophages Are Functionally Comparable With Primary Cells.
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