Cecchelli Romeo, Aday Sezin, Sevin Emmanuel, Almeida Catarina, Culot Maxime, Dehouck Lucie, Coisne Caroline, Engelhardt Britta, Dehouck Marie-Pierre, Ferreira Lino
Blood Brain Barrier Laboratory, University of Artois, Lens, France.
CNC - Center of Neurosciences and Cell Biology, University of Coimbra, Coimbra, Portugal; Biomaterials and Stem Cell-based Therapeutics Laboratory, Biocant - Center of Innovation in Biotechnology, Cantanhede, Portugal.
PLoS One. 2014 Jun 17;9(6):e99733. doi: 10.1371/journal.pone.0099733. eCollection 2014.
The human blood brain barrier (BBB) is a selective barrier formed by human brain endothelial cells (hBECs), which is important to ensure adequate neuronal function and protect the central nervous system (CNS) from disease. The development of human in vitro BBB models is thus of utmost importance for drug discovery programs related to CNS diseases. Here, we describe a method to generate a human BBB model using cord blood-derived hematopoietic stem cells. The cells were initially differentiated into ECs followed by the induction of BBB properties by co-culture with pericytes. The brain-like endothelial cells (BLECs) express tight junctions and transporters typically observed in brain endothelium and maintain expression of most in vivo BBB properties for at least 20 days. The model is very reproducible since it can be generated from stem cells isolated from different donors and in different laboratories, and could be used to predict CNS distribution of compounds in human. Finally, we provide evidence that Wnt/β-catenin signaling pathway mediates in part the BBB inductive properties of pericytes.
人血脑屏障（BBB）是由人脑内皮细胞（hBECs）形成的选择性屏障，对于确保神经元功能正常以及保护中枢神经系统（CNS）免受疾病侵害至关重要。因此，人源体外BBB模型的构建对于与CNS疾病相关的药物研发项目极为重要。在此，我们描述了一种使用脐带血来源的造血干细胞生成人BBB模型的方法。这些细胞首先分化为内皮细胞，然后通过与周细胞共培养诱导其具备BBB特性。脑样内皮细胞（BLECs）表达紧密连接和通常在脑内皮中观察到的转运蛋白，并在至少20天内维持大多数体内BBB特性的表达。该模型具有很高的可重复性，因为它可以从不同供体分离的干细胞以及在不同实验室中生成，并且可用于预测化合物在人体中的CNS分布。最后，我们提供证据表明Wnt/β-连环蛋白信号通路部分介导了周细胞的BBB诱导特性。
