Microfluidic model of the blood brain barrier

Abstract

The invention relates to culturing brain endothelial cells, and optionally astrocytes and neurons in a fluidic device under conditions whereby the cells mimic the structure and function of the blood brain barrier. Culture of such cells in a microfluidic device, whether alone or in combination with other cells, drives maturation and/or differentiation further than existing systems.

1 - 42 . (canceled) 
     
     
         43 . A method of culturing cells, comprising: a) providing a microfluidic device comprising a membrane, said membrane comprising a top surface and a bottom surface; b) coating said top surface of said membrane with laminin and said bottom surface with a mixture of collagen and fibronectin, said mixture free of laminin; c) seeding induced motor neuron progenitor cells on said top surface and brain microvascular endothelial cells on said bottom surface so as to create seeded cells; d) exposing said seeded cells to a flow of culture media for a period of time; and e) culturing said seeded cells under conditions such that said brain microvascular endothelial cells on said bottom surface form tight junctions. 
     
     
         44 . The method of  claim 43 , wherein said induced motor neuron progenitor cells are derived from induced pluripotent stem cells from a human patient diagnosed with a CNS disorder. 
     
     
         45 . The method of  claim 43 , wherein said flow promotes the differentiation of said induced motor neuron progenitor cells. 
     
     
         46 . The method of  claim 45 , wherein said induced motor neuron progenitor cells differentiate into neurons. 
     
     
         47 . The method of  claim 46 , wherein said neurons exhibit a more mature electrophysiology as compared to the same neurons cultured in a static culture 
     
     
         48 . The method of  claim 43 , wherein said induced motor neuron progenitor cells are derived from induced pluripotent stem cells from a patient diagnosed with Amyotrophic lateral sclerosis (ALS). 
     
     
         49 . The method of  claim 43 , wherein said brain microvascular endothelial cells are derived from induced pluripotent stem cells from a patient diagnosed with MCT8-specific thyroid hormone cell-membrane transporter deficiency. 
     
     
         50 . The method of  claim 43 , wherein said induced motor neuron progenitor cells were stored frozen and then thawed prior to step c). 
     
     
         51 - 80 . (canceled)