Creation of an In Vitro Brain Barrier Transport System

体外脑屏障运输系统的创建

基本信息

批准号：
7035380
负责人：
WEI ZHENG
金额：
$ 20.65万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-04-01 至 2007-03-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Brain is well protected against blood-borne xenobiotics (drugs, nutrients, and toxins) by two major barriers, i.e., blood-brain barrier (BBB) and blood-cerebrospinal fluid (CSF) barrier (BCB). The BCB, whose surface area is about one-half of the BBB, is located in brain ventricles and functions to produce CSF and transport xenobiotics between blood and CSF. We have recently established a novel immortalized choroidal epithelial cell line, named Z310 cell line. This ceil line possesses the essential morphology of the parent primary cells and, upon growing on a semipermeable membrane, forms a monolayer that restricts the free movement of paracellular leakage marker, [14C]sucrose. While the tightness of the cell monolayer, as measured by trans-epithelial electrical resistance (TEER) or paracellular leakage of [14C]sucrose, remains to be improved, we are convinced that this cell line shows a great promise as a unique in vitro blood-brain barrier transport system, as there has been no such brain cell-derived transport model in the current neurotoxicology and neuropharmacology research field. To create this novel system, we hypothesize that the tightness of the monolayer of Z310 epithelial cells can be improved by alteration of the chemical components of the culture media, by induction and promotion of tight junction assembly, and/or by genetic modulation of the expression of tight junction proteins. Thus, our specific aims are to reduce the paracellular permeability of the Z310 barrier model by modifying the culture medium components, such as using serum-free or astrocyte-conditioned culture media, to improve the tightness of the barrier structure by application of tight-junction inducing agents in cell culture medium, and to knock-in the specific gene fragments that encode the proteins or regulatory proteins associated with tight junctions in existing Z310 cells. We further design a series of experiments to validate this model system. The studies proposed in this application, if successful despite the notable risk, will lead to the creation of a novel in vitro blood-brain/CSF model for transport study of materials into brain and should have significant impact on pharmacological and toxicological investigation of CNS transport of drugs and toxicants, CNS drug development, and etiological research of brain diseases.

描述（由申请人提供）：大脑受到两个主要障碍，即血脑屏障（BBB）和脑脊液（CSF）障碍（CSF）屏障（BCB）的两个主要障碍，可以很好地保护血液传播异种生物学（药物，营养和毒素）。 BCB的表面积约为BBB的一半，位于脑室和功能中，可产生CSF并在血液和CSF之间运输异生物生物。我们最近建立了一种新颖的永生脉络膜上皮细胞系，称为Z310细胞系。该天花线具有母体原代细胞的基本形态，并且在半透明的膜上生长后，形成了限制细胞细胞泄漏标记物的自由运动的单层，[14C]蔗糖。 While the tightness of the cell monolayer, as measured by trans-epithelial electrical resistance (TEER) or paracellular leakage of [14C]sucrose, remains to be improved, we are convinced that this cell line shows a great promise as a unique in vitro blood-brain barrier transport system, as there has been no such brain cell-derived transport model in the current neurotoxicology and neuropharmacology research field.为了创建这个新型系统，我们假设可以通过改变培养基的化学成分，通过诱导和促进紧密连接组件的诱导和/或通过遗传调节紧密连接蛋白的表达来改善Z310上皮细胞单层的紧密度。因此，我们的具体目的是通过修改培养培养基组件（例如使用无血清或星形胶质细胞条件的培养基）来降低Z310屏障模型的细胞细胞渗透性，以改善屏障结构的紧密性，以通过在细胞培养基中应用紧密的蛋白质，并构成特定的蛋白质，并构成特定的蛋白质，并构成特定的蛋白质，并在内Z310细胞。我们进一步设计了一系列实验来验证该模型系统。在本应用中提出的研究（尽管有明显的风险）将导致创建一种新型的体外血脑/CSF模型，以将材料转运到大脑的运输研究，并应对药物和毒物的CNS运输的药理和毒理学研究产生重大影响。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

TWIST-1 is overexpressed in neoplastic choroid plexus epithelial cells and promotes proliferation and invasion.

DOI：
10.1158/0008-5472.can-08-3176
发表时间：
2009-03-15
期刊：
Cancer research
影响因子：
11.2
作者：
Hasselblatt M;Mertsch S;Koos B;Riesmeier B;Stegemann H;Jeibmann A;Tomm M;Schmitz N;Wrede B;Wolff JE;Zheng W;Paulus W
通讯作者：
Paulus W