Directed Differentiation of Human Pluripotent Stem Cells to Microglia.
Directed Differentiation of Human Pluripotent Stem Cells to Microglia.
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DOI：
10.1016/j.stemcr.2017.04.023
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发表时间：
2017-06-06
影响因子：
5.9
通讯作者：
Fossati V
中科院分区：
文献类型：
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作者：
Douvaras P;Sun B;Wang M;Kruglikov I;Lallos G;Zimmer M;Terrenoire C;Zhang B;Gandy S;Schadt E;Freytes DO;Noggle S;Fossati V
小胶质细胞是大脑中的免疫细胞，对中枢神经系统的正常发育和维持至关重要，并且它们在众多神经系统疾病中的参与也越来越受到认可。为了增进我们对人类小胶质细胞生物学的理解，我们设计了一种化学成分确定的方案，从多能干细胞生成人类小胶质细胞。在从胚胎干细胞和诱导多能干细胞（iPSCs）分化的30天内产生了表达CD14/CX3CR1的髓系祖细胞。这些祖细胞的进一步分化产生了具有高度活动突起的分支状小胶质细胞，表达典型的小胶质细胞标志物。基因表达和细胞因子释放分析表明，iPSC衍生的（iPSC - MG）小胶质细胞与人类原代小胶质细胞非常相似，并且与巨噬细胞有明显区别。iPSC - MG能够吞噬，并通过产生细胞内Ca2 +瞬变对ADP作出反应，而巨噬细胞则缺乏这种反应。该分化方案在多个多能干细胞系中具有高度可重复性。
从多能干细胞生成人类小胶质细胞的高效方案
iPSC衍生的小胶质细胞具有分支形态和活动突起
iPSC衍生小胶质细胞的表达和细胞因子谱与原代小胶质细胞相似
iPSC衍生的小胶质细胞能够吞噬微珠并对ADP作出反应
杜瓦拉斯及其同事通过一种化学成分确定的方案，经由髓系祖细胞从多能干细胞生成了小胶质细胞。iPSC衍生的小胶质细胞显示出高度活动的突起，能够吞噬，并通过钙瞬变对ADP作出反应。通过将多个iPSC衍生的小胶质细胞样本与原代小胶质细胞以及外周血或其他组织特异性巨噬细胞进行基因表达分析，进一步确认了小胶质细胞的特性。
Microglia, the immune cells of the brain, are crucial to proper development and maintenance of the CNS, and their involvement in numerous neurological disorders is increasingly being recognized. To improve our understanding of human microglial biology, we devised a chemically defined protocol to generate human microglia from pluripotent stem cells. Myeloid progenitors expressing CD14/CX3CR1 were generated within 30 days of differentiation from both embryonic and induced pluripotent stem cells (iPSCs). Further differentiation of the progenitors resulted in ramified microglia with highly motile processes, expressing typical microglial markers. Analyses of gene expression and cytokine release showed close similarities between iPSC-derived (iPSC-MG) and human primary microglia as well as clear distinctions from macrophages. iPSC-MG were able to phagocytose and responded to ADP by producing intracellular Ca2+ transients, whereas macrophages lacked such response. The differentiation protocol was highly reproducible across several pluripotent stem cell lines. Efficient protocol to generate human microglia from PSCs iPSC-derived microglia have ramified morphology and motile processes Expression and cytokine profiles of iPSC-derived microglia resemble primary microglia iPSC-derived microglia can phagocytose beads and respond to ADP Douvaras and colleagues generated microglia from pluripotent stem cells using a chemically defined protocol through a myeloid progenitor. iPSC-derived microglia showed highly motile processes, were able to phagocytose and responded to ADP with calcium transients. Microglial identity was further confirmed by gene expression analysis comparing multiple iPSC-derived microglia samples with primary microglia and peripheral blood or other tissue-specific macrophages.