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Evaluation of the Influence of Astrocytes on In Vitro Blood–Brain Barrier Models
Alternatives to Laboratory Animals ( IF 2.7 ) Pub Date : 2020-07-01 , DOI: 10.1177/0261192920966954
Adrián García-Salvador 1 , Alazne Domínguez-Monedero 1 , Paloma Gómez-Fernández 1 , Amaia García-Bilbao 1 , Susana Carregal-Romero 2, 3 , Joaquín Castilla 4, 5 , Felipe Goñi-de-Cerio 1
Affiliation  

In vitro blood–brain barrier (BBB) models are a useful tool to screen the permeability and toxicity of new drugs. Currently, many different in vitro BBB models coexist, but none stands out as being notably better than the rest. Therefore, there is still a need to evaluate the quality of BBB models under various conditions and assess their ability to mimic the in vivo situation. In this study, two brain endothelial cell lines (bEnd.3 and hCMEC/D3) and two epithelial-like cell lines (MDCKII and Caco-2) were selected for BBB modelling purposes. They were grown as monolayers of a single cell type, under the following conditions: in coculture with either primary or immortalised astrocytes; or in the presence of primary or immortalised astrocyte-derived conditioned media. A total of 20 different BBB models were established in this manner, in order to assess the effects of the astroglial components on the BBB phenotype in each case. To this end, six parameters were studied: the expression of selected tight junction proteins; the enzyme activities of alkaline phosphatase and of gamma glutamyl transpeptidase; the transendothelial/transepithelial electrical resistance (TEER); restriction in paracellular transport; and efflux transporter inhibition were each evaluated and correlated. The results showed that coculturing with either primary or immortalised astrocytes led to a general improvement in all parameters studied, evidencing the contribution of this cell type to effective BBB formation. Furthermore, the permeability coefficient (P e) of the tracer molecule, Lucifer Yellow, correlated with three of the six parameters studied. In addition, this study highlights the potential for the use of the Lucifer Yellow P e value as an indicator of barrier integrity in in vitro BBB models, which could be useful for screening the permeability of new drugs.

中文翻译:

星形胶质细胞对体外血脑屏障模型影响的评价

体外血脑屏障 (BBB) 模型是筛选新药渗透性和毒性的有用工具。目前,许多不同的体外 BBB 模型共存,但没有一个明显优于其他模型。因此,仍然需要评估各种条件下 BBB 模型的质量,并评估其模拟体内情况的能力。在这项研究中,选择了两种脑内皮细胞系(bEnd.3 和 hCMEC/D3)和两种上皮样细胞系(MDCKII 和 Caco-2)用于 BBB 建模目的。它们在以下条件下作为单一细胞类型的单层生长:与原代或永生化星形胶质细胞共培养;或存在原代或永生化星形胶质细胞衍生的条件培养基。这样一共建立了20个不同的BBB模型,为了评估星形胶质细胞成分对每种情况下 BBB 表型的影响。为此,研究了六个参数:选定的紧密连接蛋白的表达;碱性磷酸酶和γ谷氨酰转肽酶的酶活性;跨内皮/跨上皮电阻 (TEER);细胞旁运输的限制;和外排转运蛋白抑制分别进行评估和关联。结果表明,与原代或永生化星形胶质细胞共培养导致所有研究参数的普遍改善,证明这种细胞类型对有效 BBB 形成的贡献。此外,示踪剂分子 Lucifer Yellow 的渗透系数 (P e) 与研究的六个参数中的三个相关。此外,
更新日期:2020-07-01
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