Influence of basal media composition on barrier fidelity within human pluripotent stem cell-derived blood-brain barrier models.
Influence of basal media composition on barrier fidelity within human pluripotent stem cell-derived blood-brain barrier models.
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DOI：
10.1111/jnc.15532
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发表时间：
2021-12
影响因子：
4.7
通讯作者：
Lippmann ES
中科院分区：
文献类型：
--
作者：
Neal EH;Katdare KA;Shi Y;Marinelli NA;Hagerla KA;Lippmann ES
脑微血管内皮细胞（BMEC）是血脑屏障（BBB）的主要组成部分，对来自血流和脑的可溶性信号高度敏感。这一概念在体外延伸，其中细胞外环境也可以影响培养细胞中的BBB性质。然而，基线培养条件在多大程度上可以影响体外血脑屏障的性质仍不清楚，这对模型的变异性和再现性，以及下游的分子转运和疾病表型的评估有影响。在这里，我们通过检查人诱导多能干细胞（iPSC）衍生的BMEC样细胞在DMEM/F12和Neurobasal培养基中无血清条件下培养的BBB特性来探索这一概念，这些培养基具有完全确定的组成。我们证明了显着的差异，在被动和主动血脑屏障性能的基础媒体组合物的功能。此外，RNA测序和磷酸化蛋白质组分析揭示了各种信号通路的改变，以响应基础培养基的差异。总体而言，我们的研究结果表明，基线培养条件可以对体外BBB模型的性能产生深远的影响，在设计利用此类模型进行基础研究和临床前试验的实验时，应考虑这些影响。血脑屏障（BBB）是高度敏感的环境线索在体内，但在体外BBB模型的细胞外环境的影响还没有得到广泛的特点。使用从人类诱导多能干细胞（iPSC）分化的脑微血管内皮细胞（BMEC）样细胞，Neal，Katdare及其同事表明，不同的基础培养基成分可以显着改变被动和主动BBB特性。这些发现表明，BBB模型的用户在解释药物筛选和疾病建模测定时应注意培养基组成。这些结果也可能促使在BBB模型中使用完全定义的培养基。
It is increasingly recognized that brain microvascular endothelial cells (BMECs), the principal component of the blood-brain barrier (BBB), are highly sensitive to soluble cues from both the bloodstream and the brain. This concept extends in vitro, where the extracellular milieu can also influence BBB properties in cultured cells. However, the extent to which baseline culture conditions can affect BBB properties in vitro remains unclear, which has implications for model variability and reproducibility, as well as downstream assessments of molecular transport and disease phenotypes. Here, we explore this concept by examining BBB properties within human induced pluripotent stem cell (iPSC)-derived BMEC-like cells cultured under serum-free conditions in DMEM/F12 and Neurobasal media, which have fully defined compositions. We demonstrate notable differences in both passive and active BBB properties as a function of basal media composition. Further, RNA sequencing and phosphoproteome analyses revealed alterations to various signaling pathways in response to basal media differences. Overall, our results demonstrate that baseline culture conditions can have a profound influence on the performance of in vitro BBB models, and these effects should be considered when designing experiments that utilize such models for basic research and preclinical assays. The blood-brain barrier (BBB) is highly sensitive to environmental cues in vivo, but the influence of the extracellular milieu on in vitro BBB models has not been extensively characterized. Using brain microvascular endothelial cell (BMEC)-like cells differentiated from human induced pluripotent stem cells (iPSCs), Neal, Katdare, and colleagues show that passive and active BBB properties can be significantly altered by different basal media compositions. These findings suggest that users of BBB models should be mindful of media composition when interpreting drug screening and disease modeling assays. These results may also motivate the use of fully defined media in BBB models.
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