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A method for activating astrocytes using surface microstructures

A technology of glial cells and microstructure, applied in nervous system cells, animal cells, nervous system diseases, etc., to achieve the effect of avoiding glial scars and broad application prospects

Active Publication Date: 2020-03-31
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the existing methods of activating glial cells, such as mechanical injury, hypoxia, and stimulation of inflammatory factors, cannot avoid the formation of reactive glial scars caused by activation.

Method used

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  • A method for activating astrocytes using surface microstructures
  • A method for activating astrocytes using surface microstructures
  • A method for activating astrocytes using surface microstructures

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Using computer-aided design and wet etching methods, a regular hexagonal convex surface membrane with a side length of 30 μm was fabricated, the distance between the regular hexagons was 5 μm, and the height difference between the convex surface and the groove was 60 μm. The PDMS monomer and the initiator were mixed uniformly in the ratio of 10:1, poured on the template for reverse film, and the PDMS was cured after 30 minutes at 80 °C. Peel off the PDMS with tweezers to obtain a PDMS chip with a concave surface regular hexagonal microstructure (see figure 1 ). Put the microstructured side up into the petri dish, and fix it on the bottom of the petri dish using the stickiness of PDMS itself. Sterilize by UV irradiation overnight.

[0032] Primary extraction of astrocytes in neonatal 24h neonatal rat cerebral cortex, after removal of miscellaneous cells, the cells were grown to confluence, the cells were digested with 0.25% trypsin to prepare a cell suspension, and the...

Embodiment 2

[0034] A regular hexagonal concave surface membrane plate with a side length of 30 μm was fabricated, the distance between regular hexagons was 5 μm, and the height difference between the concave surface and the surrounding raised frame was 60 μm. PDMS chip with convex surface regular hexagonal microstructure was obtained after using PDMS reverse film. Put the microstructured side up into the petri dish, and fix it on the bottom of the petri dish using the stickiness of PDMS itself. Sterilize by UV irradiation overnight.

[0035] The primary extracted neonatal mouse astrocyte suspension was added to the surface of the PDMS microstructure and placed in an incubator for adherent culture for 1-14 days. Glial cells grown on the convex surface were identified, and the results showed that the cells underwent changes similar to those on the concave surface, and the intracellular expression of vimentin and stemness-related protein SOX-2 were increased ( Figure 5 ).

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Abstract

The invention provides a method for activating astrocytes by using surface microstructures, which belongs to the field of cell culture technology. The specific steps are: (1) obtaining a PDMS cell culture surface with uneven microstructures; The subcultured glial cells are the activation objects. After the cells grow to the cell fusion, the cells are digested with 0.25% trypsin to make a cell suspension. (3) Add the cell suspension to the surface of the microstructure. After the cells adhere to the wall, Cultured for 1-14 days; (4) The astrocytes obtained by the above method were identified to show a series of activation performances such as EMT transformation, enhanced cell stemness, and enhanced glucose transport. The advantage of the present invention is that glial cells can be activated by using the physical microstructure on the surface of the culture, and glial scars caused by reactive hyperplasia of glial cells can be avoided. The invention has broad application prospects in the treatment of nervous system damage and degenerative diseases.

Description

technical field [0001] The invention relates to cell culture technology, and particularly provides a method for activating astrocytes by utilizing surface microstructures. Background technique [0002] Astrocytes are the largest, most numerous and most widely distributed glial cells in the central nervous system, with a large number of radial processes. Astrocytes play a decisive role in neural development, synapse formation, blood-brain barrier construction, energy metabolism, etc., and are essential for maintaining brain homeostasis and normal neuron function. [0003] Astrocytes exist in two states: resting state and activated state. When the nervous system is stimulated by various harmful factors, such as hypoxia, trauma, inflammation, etc., astrocytes transform from a resting state to an activated state and become activated astrocytes. The role of activated glial cells is a "double-edged sword". On the one hand, activated glial cells rapidly migrate and surround the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/079A61K35/30A61P25/00A61P9/10A61P25/16A61P25/28
Inventor 秦建华张敏张旭许慧
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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