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Method utilizing surface microstructure to activate neural astrocyte

A glial cell and microstructure technology, applied in nervous system cells, animal cells, nervous system diseases, etc., to achieve broad application prospects and avoid glial scars

Active Publication Date: 2016-06-08
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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  • Method utilizing surface microstructure to activate neural astrocyte
  • Method utilizing surface microstructure to activate neural astrocyte
  • Method utilizing surface microstructure to activate neural astrocyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Using computer-aided design and wet etching method, a regular hexagonal convex surface membrane plate 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. Mix the PDMS monomer and initiator evenly at a ratio of 10:1, pour it on the template for reverse filming, and cure the PDMS after 30 minutes at 80°C. Use tweezers to peel off the PDMS to obtain a PDMS chip with a regular hexagonal microstructure on the concave surface (see figure 1 ). Put the side with the microstructure up into the Petri dish, and use the viscosity of PDMS itself to fix it to the bottom of the Petri dish. Sterilize overnight by UV irradiation.

[0032] Primary extraction of newborn 24h neonatal mouse cerebral cortex astroglial cells, after removal of miscellaneous cells, the cells were grown to confluence, the cells were digested with 0.25% trypsin to make a cell suspension, an...

Embodiment 2

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

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

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Abstract

The invention provides a method utilizing a surface microstructure to activate neural astrocyte, and belongs to the technical field of cell culture. The method comprises the following steps: (1) obtaining a PDMS cell culture surface with an uneven microstructure; (2) adopting primarily cultured neurogliocyte as the activation object, and after the cells grow to the phase of cell fusion, digesting cells by pancreatin (0.25%) to prepare cell suspension; (3) adding the cell suspension to the microstructure surface, and after cell attachment, culturing the cells for 1 to 14 days; (4) identifying the neural astrocyte obtained by the method mentioned above to find a series of activation expressions that cells carry out EMT conversion, cell stem property is enhanced, and glucose transportation is strengthened. The method has the advantage that a physical microstructure on the surface of a culture medium is utilized to activate neural astrocyte and at the same time, formation of glial scar caused by reactive hyperplasia of colloid cells can be avoided. The provided method has a wide application prospect in the treatment of destructive disease and degenerative disease of nervous system.

Description

technical field [0001] The invention relates to cell culture technology, and particularly provides a method for activating astrocytes by using 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 many aspects such as neural development, synapse formation, blood-brain barrier construction, energy metabolism, etc., and are crucial for maintaining brain homeostasis and normal function of neurons. [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 migrate rapid...

Claims

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Application Information

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Patent Type & Authority Applications(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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