Generation and Functional Characterization of Monocytes and Macrophages Derived from Human Induced Pluripotent Stem Cells

Q2 Biochemistry, Genetics and Molecular Biology

Current Protocols in Stem Cell Biology Pub Date : 2020-03-11 DOI:10.1002/cpsc.108

Xu Cao, Francijna E. van den Hil, Christine L. Mummery, Valeria V. Orlova

{"title":"Generation and Functional Characterization of Monocytes and Macrophages Derived from Human Induced Pluripotent Stem Cells","authors":"Xu Cao, Francijna E. van den Hil, Christine L. Mummery, Valeria V. Orlova","doi":"10.1002/cpsc.108","DOIUrl":null,"url":null,"abstract":"Monocytes and macrophages are essential for immune defense and tissue hemostasis. They are also the underlying trigger of many diseases. The availability of robust and short protocols to induce monocytes and macrophages from human induced pluripotent stem cells (hiPSCs) will benefit many applications of immune cells in biomedical research. Here, we describe a protocol to derive and functionally characterize these cells. Large numbers of hiPSC-derived monocytes (hiPSC-mono) could be generated in just 15 days. These monocytes were fully functional after cryopreservation and could be polarized to M1 and M2 macrophage subtypes. hiPSC-derived macrophages (iPSDMs) showed high phagocytotic uptake of bacteria, apoptotic cells, and tumor cells. The protocol was effective across multiple hiPSC lines. In summary, we developed a robust protocol to generate hiPSC-mono and iPSDMs which showed phenotypic features of macrophages and functional maturity in different bioassays. © 2020 The Authors.Basic Protocol 1: Differentiation of hiPSCs toward monocytesSupport Protocol 1: Isolation and cryopreservation of monocytesSupport Protocol 2: Characterization of monocytesBasic Protocol 2: Differentiation of different subtypes of macrophagesSupport Protocol 3: Characterization of hiPSC-derived macrophages (iPSDMs)Support Protocol 4: Functional characterization of different subtypes of macrophages","PeriodicalId":53703,"journal":{"name":"Current Protocols in Stem Cell Biology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2020-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpsc.108","citationCount":"18","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Stem Cell Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpsc.108","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}

引用次数: 18

Abstract

Monocytes and macrophages are essential for immune defense and tissue hemostasis. They are also the underlying trigger of many diseases. The availability of robust and short protocols to induce monocytes and macrophages from human induced pluripotent stem cells (hiPSCs) will benefit many applications of immune cells in biomedical research. Here, we describe a protocol to derive and functionally characterize these cells. Large numbers of hiPSC-derived monocytes (hiPSC-mono) could be generated in just 15 days. These monocytes were fully functional after cryopreservation and could be polarized to M1 and M2 macrophage subtypes. hiPSC-derived macrophages (iPSDMs) showed high phagocytotic uptake of bacteria, apoptotic cells, and tumor cells. The protocol was effective across multiple hiPSC lines. In summary, we developed a robust protocol to generate hiPSC-mono and iPSDMs which showed phenotypic features of macrophages and functional maturity in different bioassays. © 2020 The Authors.

Basic Protocol 1: Differentiation of hiPSCs toward monocytes

Support Protocol 1: Isolation and cryopreservation of monocytes

Support Protocol 2: Characterization of monocytes

Basic Protocol 2: Differentiation of different subtypes of macrophages

Support Protocol 3: Characterization of hiPSC-derived macrophages (iPSDMs)

Support Protocol 4: Functional characterization of different subtypes of macrophages

Abstract Image

查看原文本刊更多论文

人诱导多能干细胞单核细胞和巨噬细胞的生成及功能表征

单核细胞和巨噬细胞对免疫防御和组织止血至关重要。它们也是许多疾病的潜在诱因。从人诱导多能干细胞(hiPSCs)中诱导单核细胞和巨噬细胞的强大和短的方案的可用性将有利于免疫细胞在生物医学研究中的许多应用。在这里，我们描述了一个方案来推导和功能表征这些细胞。大量hipsc衍生单核细胞(hiPSC-mono)可以在短短15天内生成。这些单核细胞冷冻保存后功能完全，可分化为M1和M2巨噬细胞亚型。hipsc衍生的巨噬细胞(iPSDMs)对细菌、凋亡细胞和肿瘤细胞表现出高度的吞噬摄取。该方案在多个hiPSC系中有效。总之，我们开发了一种强大的方案来生成hiPSC-mono和iPSDMs，它们在不同的生物测定中显示巨噬细胞的表型特征和功能成熟度。©2020作者。基本方案1:hipsc向单核细胞的分化支持方案1:单核细胞的分离和冷冻保存支持方案2:单核细胞的表征基本方案2:不同亚型巨噬细胞的分化支持方案3:hipsc衍生巨噬细胞(iPSDMs)的表征支持方案4:不同亚型巨噬细胞的功能表征

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Current Protocols in Stem Cell Biology Biochemistry, Genetics and Molecular Biology-Cell Biology

自引率

0.00%

发文量

期刊介绍： Published in affiliation with the International Society for Stem Cell Research (ISSCR), Current Protocols in Stem Cell Biology (CPSC) covers the most fundamental protocols and methods in the rapidly growing field of stem cell biology. Updated monthly, CPSC will constantly evolve with thelatest developments and breakthroughs in the field. Drawing on the expertise of leading researchers from around the world, Current Protocols in Stem Cell Biology includes methods and insights that will enhance the progress of global research.