A Simplified, Fully Defined Differentiation Scheme for Producing Blood-Brain Barrier Endothelial Cells from Human iPSCs
A Simplified, Fully Defined Differentiation Scheme for Producing Blood-Brain Barrier Endothelial Cells from Human iPSCs
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DOI：
10.1016/j.stemcr.2019.05.008
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发表时间：
2019-06-11
影响因子：
5.9
通讯作者：
Lippmann, Ethan S.
中科院分区：
文献类型：
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作者：
Neal, Emma H.;Marinelli, Nicholas A.;Lippmann, Ethan S.
人类诱导多能干细胞(IPSC)来源的发育谱系是体外机械询问、药物发现和疾病建模的关键工具。IPSCs已经被分化为具有血脑屏障(BBB)特性的内皮细胞，其特征是高跨内皮电阻(TEER)、低被动通透性和主动转运蛋白功能。典型的协议使用未定义的组件，这给区分过程带来了不可接受的可变性。我们证明，用完全定义的成分替换血清，从普通的介质补充剂到胰岛素、转铁蛋白和硒的简单混合物，在多个IPSC系中产生TEER在2,000-8,000欧加×厘米(2)范围内的BBB内皮，具有适当的标记表达和活跃的转运蛋白。全定义培养液的使用极大地提高了分化的一致性，BBB内皮与IPSC来源的星形胶质细胞共培养产生了一个强大的体外神经血管模型。这种明确的分化方案应该能够广泛地使人血脑屏障内皮细胞用于不同的应用。
Human induced pluripotent stem cell (iPSC)-derived developmental lineages are key tools for in vitro mechanistic interrogations, drug discovery, and disease modeling. iPSCs have previously been differentiated to endothelial cells with blood-brain barrier (BBB) properties, as defined by high transendothelial electrical resistance (TEER), low passive permeability, and active transporter functions. Typical protocols use undefined components, which impart unacceptable variability on the differentiation process. We demonstrate that replacement of serum with fully defined components, from common medium supplements to a simple mixture of insulin, transferrin, and selenium, yields BBB endothelium with TEER in the range of 2,000-8,000 Omega x cm(2) across multiple iPSC lines, with appropriate marker expression and active transporters. The use of a fully defined medium vastly improves the consistency of differentiation, and co-culture of BBB endothelium with iPSC-derived astrocytes produces a robust in vitro neurovascular model. This defined differentiation scheme should broadly enable the use of human BBB endothelium for diverse applications.