Development of spinal cord on a microfluidic chip
Inventors
Kerns, S. Jordan • Wen, Norman • Lucchesi, Carolina • Hinojosa, Christopher David • Fraser, Jacob • Hamilton, Geraldine • Vatine, Gad • Sances, Samuel • Svendsen, Clive • Levner, Daniel • Sareen, Dhruv
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12091650-B2
Publication Date
2024-09-17
Expiration Date
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.
Core Innovation
The invention relates to a microfluidic dual microchannel system for culturing cells in which a first microchannel and a second microchannel are separated by a membrane. Human ventral spinal neuron progenitor cells are seeded in the first microchannel, while brain microvascular endothelial cells are seeded in the second microchannel. The system supports culturing with a flowing culture media and enables direct physical contact between at least a portion of the ventral spinal neuron progenitor cells and the brain microvascular endothelial cells through the membrane.
In one aspect, human ventral spinal neuron progenitor cells are differentiated with a flowing culture media in the first microchannel, and at least a portion of the cells are cultured to express at least one marker of a spinal motor neuron. The patent further characterizes cultured cells using marker proteins associated with spinal motor neuron identity. In related embodiments, induced pluripotent human stem cells are differentiated to neural ectoderm, and the neural ectoderm is exposed to retinoic acid and a sonic hedgehog agonist to pattern ventral spinal neural progenitor cells.
In additional embodiments, the seeded cell set includes glial cells seeded within the first microchannel, with brain microvascular endothelial cells seeded within the second microchannel so that the co-cultured populations are in direct physical contact through the membrane. The invention recreates a direct neuron-endothelial interaction environment within a dual microchannel microfluidic device while culturing patterned ventral spinal neural progenitor cells under flowing culture media.
Claims Coverage
The document provides three independent methods of culturing cells. Across the independent claims, the inventive features focus on a dual microchannel microfluidic device separated by a membrane, co-seeding ventral spinal neuron progenitor cells and brain microvascular endothelial cells into separate microchannels, and differentiating and/or patterning the neuron progenitors under flowing culture media to achieve direct physical contact through the membrane, with optional glial co-seeding and marker-based characterization.
Differentiating ventral spinal neuron progenitors with flowing media in a dual microchannel device separated by a membrane
A dual microchannel microfluidic device comprising a first microchannel and a second microchannel separated by a membrane; human ventral spinal neuron progenitor cells seeded within said first channel; brain microvascular endothelial cells seeded within said second microchannel; differentiating at least a portion of said human ventral spinal neuron progenitor cells with a flowing culture media in said first microchannel; and culturing said human ventral spinal neuron progenitor cells to express at least one marker of a spinal motor neuron.
Patterning ventral spinal neural progenitor cells and creating direct physical contact through the membrane under flowing culture
Providing induced pluripotent human stem cells, brain microvascular endothelial cells, and a dual microchannel microfluidic device comprising a first microchannel and second microchannel separated by a membrane; differentiating said induced pluripotent stem cells to neural ectoderm; exposing said neural ectoderm to retinoic acid and sonic hedgehog agonist to pattern ventral spinal neural progenitor cells; seeding said ventral spinal neural progenitor cells in said first microchannel on said membrane and seeding said brain microvascular endothelial cells in said second microchannel on said membrane of said microfluidic device so as to create seeded cells; and culturing said ventral spinal neural progenitor cells with a flowing culture media such that at least a portion of said patterned ventral spinal neural progenitor cells and said brain microvascular endothelial cells are in direct physical contact with each other through said membrane.
Differentiating ventral spinal neuron progenitors with flowing media while co-seeding glial cells for direct physical contact through the membrane
Providing human ventral spinal neuron progenitor cells, brain microvascular endothelial cells, glial cells and a dual microchannel microfluidic device comprising a first microchannel and a second microchannel separated by a membrane; seeding said human ventral spinal neuron progenitor cells and said glial cells within said first microchannel and seeding said brain microvascular endothelial cells within said second microchannel so as to create seeded cells; and differentiating said seeded human ventral spinal neuron progenitor cells with a culture media flowing over said human ventral spinal neuron progenitor cells such that at least a portion of said human ventral spinal neuron progenitor cells and brain microvascular endothelial cells are in direct physical contact with each other.
Across the independent claims, the core coverage is a dual microchannel microfluidic device separated by a membrane that enables co-culture of ventral spinal neuron progenitor cells and brain microvascular endothelial cells under flowing culture media. The claims further require differentiation and/or patterning of ventral spinal neural progenitor cells, including retinoic acid and sonic hedgehog agonist exposure when using induced pluripotent human stem cells, and require that at least a portion of the neuron progenitors and endothelial cells are in direct physical contact through the membrane, with optional glial co-seeding and marker expression constraints.
Stated Advantages
Documented Applications
No documented applications found
Interested in licensing this patent?