Blood-Brain Barrier in vitro Model: A Tissue Engineering Approach and Validation

Authors

Zhiqi Zhang, Florida International UniversityFollow

Document Type

Dissertation

Major/Program

Biomedical Engineering

First Advisor's Name

Chenzhong Li

First Advisor's Committee Title

Committee Chair

Second Advisor's Name

Ophelia Weeks

Third Advisor's Name

Nikolaos Tsoukias

Fourth Advisor's Name

Wei-Chiang Lin

Fifth Advisor's Name

Yen-Chih Huang

Keywords

bood-brain barrier, tissue engineering, immortalized cell lines, in vitro modeling

Date of Defense

7-7-2010

Abstract

This dissertation evaluated the feasibility of using commercially available immortalized cell lines in building a tissue engineered in vitro blood-brain barrier (BBB) co-culture model for preliminary drug development studies. Mouse endothelial cell line and rat astrocyte cell lines purchased from American Type Culture Collections (ATCC) were the building blocks of the co-culture model. An astrocyte derived acellular extracellular matrix (aECM) was introduced in the co-culture model to provide a novel in vitro biomimetic basement membrane for the endothelial cells to form endothelial tight junctions. Trans-endothelial electrical resistance (TEER) and solute mass transport studies were engaged to quantitatively evaluate the tight junction formation on the in-vitro BBB models. Immuno-fluorescence microscopy and Western Blot analysis were used to qualitatively verify the in vitro expression of occludin, one of the earliest discovered tight junction proteins. Experimental data from a total of 12 experiments conclusively showed that the novel BBB in vitro co-culture model with the astrocyte derived aECM (CO+aECM) was promising in terms of establishing tight junction formation represented by TEER values, transport profiles and tight junction protein expression when compared with traditional co-culture (CO) model setups and endothelial cells cultured alone. Experimental data were also found to be comparable with several existing in vitro BBB models built from various methods. In vitro colorimetric sulforhodamine B (SRB) assay revealed that the co-cultured samples with aECM resulted in less cell loss on the basal sides of the insert membranes than that from traditional co-culture samples. The novel tissue engineering approach using immortalized cell lines with the addition of aECM was proven to be a relevant alternative to the traditional BBB in vitro modeling.

Identifier

FI10081003

Recommended Citation

Zhang, Zhiqi, "Blood-Brain Barrier in vitro Model: A Tissue Engineering Approach and Validation" (2010). FIU Electronic Theses and Dissertations. 246.
https://digitalcommons.fiu.edu/etd/246