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Three-channel microfluidic chip for establishing three kinds of cell in-vitro co-culture models

A microfluidic chip, cell body technology, applied in the field of biomedical engineering, can solve problems such as detection difficulties

Inactive Publication Date: 2016-04-06
SHENZHEN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, due to the limitation of the structure of the transwell chamber, the above three types of cells can only realize two kinds of direct contact in the three-dimensional space.
The current on-chip detection of O 2 Concentration and partial pressure of oxygen have certain difficulties
[0005] Therefore, by using a microfluidic chip, the NVU model was constructed in vitro from neurons, astrocytes, and brain microvascular endothelial cells, and then chemical hypoxia was applied to construct Na 2 S 2 o 4 remove O from the medium 2 , simulating chemical glucose and oxygen deprivation injury, and establishing an in vitro stroke model is still blank

Method used

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Experimental program
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Effect test

Embodiment Construction

[0027] As mentioned above, the structure of the three-channel microfluidic chip used to establish the in vitro co-culture model of three types of cells includes three flow channels that can be inoculated with different cells and two through-flow channels designed to realize the communication of different cells. Since astrocytes act as a bridge in the vascular nerve unit and are responsible for the important role of communicating neurons and brain microvascular endothelial cells, astrocytes are seeded on the middle channel. Fabrication of three-channel microfluidic chip SU8 template and PDMS chip:

[0028] 1) Use a 3-inch silicon wafer with a clean surface as the substrate, pour 1mL of SU8-3010 on it, and complete the coating on the glue spreader at 3500rmp-30 seconds (the structure height is 10 microns);

[0029] 2) Bake on a hot plate at 65 degrees for 1 minute, turn to 95 degrees and continue baking for 3 minutes, and use mask 1 to lithographically penetrate the fine channel...

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Abstract

The invention provides a three-channel microfluidic chip for establishing three kinds of cell in-vitro co-culture models. The three-channel microfluidic chip is characterized by comprising three flow channels capable of being used for inoculating different element cells respectively, and tiny flow channels which are designed for communication among different cells, and connected between each two flow channels in a communication manner; astrocytes in a nervus vascularis unit are taken as a bridge, and role the important role of communicating nerve cells and cerebral microvascular endothelial cells, therefore, the astrocytes are inoculated in the middle flow channel. The three-channel microfluidic chip has the advantages that (1) the nervus vascularis unit models established by the three-channel microfluidic chip can truly show the real framework in the brain of which the astrocytes are taken as the bridge and are connected with the other two element cells together, namely, nerve cells-astrocytes-cerebral microvascular endothelial cells; and (2) the three-channel microfluidic chip can represent the O2 of different concentrations, so to as further reflect the ischemic hypoxemia states of the three element cells in the nervus vascularis unit after the oxygen-glucose deprivation damage.

Description

technical field [0001] The invention relates to a three-channel microfluidic chip applied to a three-cell co-culture model of a neurovascular unit after chemical glucose and oxygen deprivation injury, and belongs to the technical field of biomedical engineering. Background technique [0002] The neurovascular unit (neurovascular unit, NVU) maintains the normal physiological functions of the nervous system, and its basic unit is composed of neurons-glial cells-vascular, including neurons, vascular endothelial cells, astrocytes, and microglia. , pericytes and other functional cell types and extracellular matrix [1] . Mechanisms such as cell-to-cell signaling and neurovascular coupling among the components of NVU are the structural basis for ensuring neuron function and normal cerebral blood flow, and a variety of neurotransmitters, protein molecules, ions, and media are also involved . Among them, astrocytes communicate with neurons through cell protrusions, and at the same...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12N5/0793C12N5/079C12N5/071
CPCC12M23/16C12N5/0602C12N5/0619C12N5/0622C12N2502/086
Inventor 褚晓凡陈艳罗翰舒伟良王垚磊
Owner SHENZHEN PEOPLES HOSPITAL
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