A Quasi-Physiological Microfluidic Blood-Brain Barrier Model for Brain Permeability Studies.
A Quasi-Physiological Microfluidic Blood-Brain Barrier Model for Brain Permeability Studies.
复制标题
DOI：
10.3390/pharmaceutics13091474
复制
发表时间：
2021-09-15
期刊：
影响因子：
5.4
通讯作者：
Cucullo L
中科院分区：
文献类型：
--
作者：
Noorani B;Bhalerao A;Raut S;Nozohouri E;Bickel U;Cucullo L
基于微流体的器官芯片技术允许开发一类新的体外血脑屏障（BBB）模型，其概括了传统二维平台无法实现的脑微血管的许多血液动力学和结构特征。在这里，我们描述并验证了一种新的微流控血脑屏障模型，密切模仿原位。将诱导多能干细胞（iPSC）衍生的脑微血管内皮细胞（BMEC）与原代人周细胞和星形胶质细胞在共培养物中并置，以实现BBB特异性特征，例如低细胞旁渗透性、流出活性和渗透反应。使用高灵敏度LC-MS/MS程序评估[13 C12]蔗糖和[13 C6]甘露醇的渗透系数。由此产生的血脑屏障显示连续的紧密连接模式，低渗透性甘露醇和蔗糖，和准生理反应高渗开放和p-糖蛋白抑制剂治疗，如证明血脑屏障完整性下降和罗丹明123的渗透性增加，分别。血管通道近腔侧的星形胶质细胞和周细胞提供了形成紧密屏障所需的环境线索，并延长了模型的长期生存能力，用于时程研究。总之，我们的新型多培养微流体平台展示了复制准生理脑微血管的能力，从而能够开发高度预测性和诊断相关的BBB模型。
Microfluidics-based organ-on-a-chip technology allows for developing a new class of in-vitro blood-brain barrier (BBB) models that recapitulate many hemodynamic and architectural features of the brain microvasculature not attainable with conventional two-dimensional platforms. Herein, we describe and validate a novel microfluidic BBB model that closely mimics the one in situ. Induced pluripotent stem cell (iPSC)-derived brain microvascular endothelial cells (BMECs) were juxtaposed with primary human pericytes and astrocytes in a co-culture to enable BBB-specific characteristics, such as low paracellular permeability, efflux activity, and osmotic responses. The permeability coefficients of [13C12] sucrose and [13C6] mannitol were assessed using a highly sensitive LC-MS/MS procedure. The resulting BBB displayed continuous tight-junction patterns, low permeability to mannitol and sucrose, and quasi-physiological responses to hyperosmolar opening and p-glycoprotein inhibitor treatment, as demonstrated by decreased BBB integrity and increased permeability of rhodamine 123, respectively. Astrocytes and pericytes on the abluminal side of the vascular channel provided the environmental cues necessary to form a tight barrier and extend the model’s long-term viability for time-course studies. In conclusion, our novel multi-culture microfluidic platform showcased the ability to replicate a quasi-physiological brain microvascular, thus enabling the development of a highly predictive and translationally relevant BBB model.