Direct-Contact BBB Model via Astrocyte Lawn
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Solution Overview
Problem
Current in vitro models for the blood-brain barrier (BBB) lack a physiologically representative configuration due to indirect interactions between astrocytes, pericytes, and endothelial cells, limiting their ability to accurately model drug permeation and transport across the BBB.
Innovation Solution
A direct-contact coculture and triculture system where brain microvessel endothelial cells (BMECs) are plated on a pre-formed lawn of astrocytes and pericytes on the apical surface of a culture chamber, allowing for optimal interaction and mimicking the in vivo environment of the BBB.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If indirect coculture configuration is used with Transwell filter support, then ease of manufacture is improved, but physiological relevance and cell-cell interaction quality deteriorate
Solution Approach 1:
The patent removes the Transwell filter support from the coculture system, extracting the barrier that prevented direct cell-cell contact. By eliminating this intermediate structure, BMECs can directly interact with astrocytes and pericytes, achieving physiological relevance while maintaining manufacturing simplicity through a straightforward well plate configuration
Solution Approach 2:
The patent eliminates the Transwell filter as an intermediary structure that mediated but limited cell interactions. By removing this mediator, the system allows direct physiological interactions between BBB components, improving model accuracy while the well plate provides sufficient structural support without interfering with cell behavior
2Device complexity
If monoculture configuration is used, then device complexity is reduced, but ability to model drug permeation accurately deteriorates
Solution Approach 1:
The patent merges multiple cell types (BMECs, astrocytes, and pericytes) into a single integrated coculture system within one well plate. This combination creates a comprehensive BBB model that accurately predicts drug permeation by incorporating all relevant cellular components and their interactions, while the unified configuration keeps the system relatively simple
3Ease of operation
If indirect triculture system is used with filter support, then ease of operation is improved, but extent of cell coverage and connection quality deteriorate
Solution Approach 1:
The patent removes the Transwell filter support that limited cell coverage area. By extracting this physical barrier, cells can spread and form continuous monolayers across the entire well plate surface, achieving complete coverage and extensive cell-cell connections while the protocol remains straightforward to execute
Data Source
AI summary
The present disclosure generally relates to a process to prepare a cell culture system that mimics the structure of blood brain barrier (BBB) and are useful to study the functions thereof. In particular, the present invention relates to a direct-contact coculture and triculture systems prepared by plating BMECs on a pre-formed lawn of coculture of astrocytes and pericytes on the apical surface of a culture-chamber to achieve a truly direct contact triculture model for BBB. The cell culture systems disclosed herein are also useful for studying the functions of the blood brain barrier and predicting the efficacy and potential toxicity of a drug candidate.