摘要
目的比较不同类型体外血脑屏障模型(blood brain barrier model,BBBM)的建立及其屏障功能特点,为构建血脑屏障(BBB)毒性筛检模型提供选择依据。方法以Wistar乳鼠脑毛细血管内皮细胞(brain capillary endothelial cells,BCECs)和星型胶质细胞(astrocytes,As)共培养构建BBBM;以渗漏试验、跨内皮细胞阻抗(transendothelial electrical resistance,TEER)测量和荧光黄(lucifer yellow,LY)通透性试验评判模型的屏障功能。结果建立分别与12孔和24孔板相匹配的各4种BBBM:单细胞模型、接触模型、非接触模型和间接模型。镜下细胞融合完整的模型,渗漏试验结果呈阳性;各模型TEER于细胞融合完整后的2～4 d到达峰值。其中以接触模型和非接触模型的TEER最为突出,12孔、24孔接触模型TEER值分别为(241±4.5)、(297±5.6)Ω/cm2;12孔、24孔非接触模型TEER值分别为(233±5.6)、(273±4.6)Ω/cm2。4种模型中,12孔接触模型和非接触模型具有较低的通透系数(permeability coefficient,Pe),其值分别是(0.92±0.07)×10-3cm/min和(1.06±0.05)×10-3cm/min。结论 12孔BBB非接触模型的建立周期短、屏障特征明显,且易于实施通透性测试操作,可作为构建BBB毒性筛检模型的优先选择。
Objective To compare the establishment and barrier traits of different in vitro blood brain barrier models （BBBMs） to supply basis for further construction of BBB toxicity screening models. Methods BBBMs were established through the co-culture of brain capillary endothelial cells （BCECs） and astroeytes （As）. 4-hour leaking test, transendothelial electrical resistance （TEER） measurement and Lucifer Yellow （LY） permeability test were performed to evaluate the barrier function of BBBMs. Results BBBMs including BCECs monolayer model, contact model, non-contact model and 1/2As-conditioned medium indirect model which were matched with 24-well and 12-well plate separately were established. 4-hour leaking test showed positive in these BBBMs ,the BCECs of which fused entirely. 2 to 4 days after BCECs＇ complete confluence, TEER values of all models reached the peak. Among these, TEERs of contact model and non-contact model were the most outstanding, which were （241±4.5） and （297±5.6） ）/cm2 in 12-well and 24-well contact model, and （233±5.6） and （273±4.6） Ω/cm2 in 12-well and 24-well non-contact model respectively. Meanwhile, in 4 kinds of BBBMs, 12-well contact model and 12-well non-contact model had lower permeability coefficient （Pc）, which were （0.92±0.07）×10-3 cndmin and （1.06±0.05）×10-3 cm/min respectively. Conclusion For the advantages of short preparation time, promising BBB characteristics and easy to implement permeability test, 12-well non-contact BBBM should be the first choice for constructing BBB toxicity screening models.
出处
《环境与健康杂志》
CAS
CSCD
北大核心
2012年第2期127-130,F0003,共5页
Journal of Environment and Health
基金
国家自然科学基金(81072260)
关键词
血脑屏障模型
屏障功能
通透系数
Blood brain barrier model
Barrier functions
Permeability coefficient
