Neuronal and neuroglial cell ordered co-culture device and preparation method thereof, and neuronal and neuroglial cell ordered co-culture method

A glial cell and neuron technology, applied in the field of cell biology, can solve the problems of cell floating, short cell survival time, and inability to ensure that neurons and glial cells are in a good time period for cultivation at the same time.

Inactive Publication Date: 2014-11-05
民航总医院 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many deficiencies when this device is used for orderly co-cultivation of neurons and glial cells. First, the cells are seeded in the microfluidic channels, and the cells are unevenly distributed in the narrow and long channels; Due to the small amount of medium in the channel, the survival time of the cells is very short, and a short period of time cannot ensure that the neurons and glial cells are cultured at the same time; if the medium is constantly replaced, it is easy to make the cells float. Ultimately unstable death; third, the cell capacity in the microfluidic channel is small, and the group effect used to study the interaction between the two cells is not obvious, which is not conducive to observation and research
In addition, because neuron cells cannot be subcultured, and the above-mentioned device requires that the intervals between inoculation of various cells should not be too long, the above-mentioned device is difficult to be used for the study of the interaction between neurons and glial cells

Method used

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  • Neuronal and neuroglial cell ordered co-culture device and preparation method thereof, and neuronal and neuroglial cell ordered co-culture method
  • Neuronal and neuroglial cell ordered co-culture device and preparation method thereof, and neuronal and neuroglial cell ordered co-culture method

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Experimental program
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Effect test

Embodiment 1

[0066] 1) Take a 3*3 cm cover glass, first soak the surface in a solution of fecivic acid (conc. Milliliter of 0.1% poly-lysine, incubate for 1.0 hour, absorb the remaining solution, wash 3 times with phosphate buffer (pH 7.4), dry in the air, and set aside;

[0067]2) Using micromachining technology, prepare a group of concave linear microstructure units on a plexiglass plate, the concave linear microstructure units are composed of two grooves arranged in sequence from left to right, wherein the left The cross section of the side groove is rectangular, the length is 1.0 cm, and the width is 0.5 cm; the cross section of the right groove is rectangular, the length is 1.0 cm, and the width is 1.0 cm; the left groove and the right groove The distance between them is 2000 microns;

[0068] 3) The plexiglass plate with one group of concave linear microstructure units obtained in step 2) is turned over twice with polydimethylsiloxane to obtain a polydimethylsiloxane template, the p...

Embodiment 2

[0077] 1) Take a cell culture dish with a diameter of 25 mm, add 1.0 ml of 0.1% L-polylysine dropwise on the bottom surface, incubate for 1.0 hour, absorb the remaining solution, and wash with phosphate buffer (pH7.4) 3 times, dry and set aside;

[0078] 2) Using micromachining technology, prepare at least one group of concave linear microstructure units on a plexiglass plate, the concave linear microstructure units are composed of three grooves arranged in sequence from left to right; wherein, the The cross section of the groove on the left side is rectangular, with a length of 1.8 cm and a width of 0.5 cm; the cross section of the middle groove is rectangular, with a length of 1.8 cm and a width of 1.1 cm; the cross section of the right groove is It is rectangular, with a length of 1.8 cm and a width of 0.5 cm; the distance between the left groove and the middle groove, and the distance between the middle groove and the right groove is 1000 microns;

[0079] 3) The plexigla...

Embodiment 3

[0088] 1) Take a cell culture dish with a diameter of 25 mm, add 1.0 ml of 0.1% poly-lysine dropwise on the bottom surface, incubate for 1.0 hour, absorb the remaining solution, and wash 3 times with phosphate buffer (pH7.4) , dry and set aside;

[0089] 2) Using micromachining technology, prepare at least one group of concave linear microstructure units on the plexiglass plate, the concave linear microstructure units are composed of three grooves arranged in sequence from left to right; wherein, the left The cross section of side groove is rectangular, and length is 1.8 centimeters, and width is 0.8 centimeter; The cross section of described middle groove is rectangular, and length is 1.8 centimeters, and width is 0.5 centimeter; The cross section of described right side groove is Rectangular with a length of 1.8 cm and a width of 0.8 cm; the distance between the left groove and the middle groove, and the middle groove and the right groove are both 500 μm;

[0090] 3) The pl...

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Abstract

The invention relates to the cytobiology field, and discloses a neuronal cell and neuroglial cell ordered co-culture device and a preparation method and an application thereof. The device includes a substrate and a polydimethylsiloxane stamper, wherein the polydimethylsiloxane stamper can be removably compounded on the substrate; the polydimethylsiloxane stamper includes at least one micropore unit, the micropore unit includes at least one first through hole perpendicular to the substrate and at least one second through hole perpendicular to the substrate, the first through hole and the second through hole are arranged successively, and the first through hole and the second through hole respectively with the surface of the substrate form grooves having one end with an opening; the distance between the first through hole and the second through hole is 500 [mu]m-2000 [mu]m. Neuronal cells and neuroglial cells are inoculated through the first through hole and the second through hole which are perpendicular to the substrate, uniformity of the two kinds of cells is easy to control during inoculation in regions, cell quantity which can be used is increased, a population effect is obvious, and observation and study are facilitated.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a device for orderly co-cultivation of neurons and glial cells, a preparation method thereof and a method for orderly co-cultivation of neurons and glial cells. Background technique [0002] Research results in recent years have shown that there is not only material exchange between neurons and glial cells, but also information exchange. How does the interaction mode between neurons and glial cells affect their material and information exchange, and the correlation between changes in their interaction mode and various brain diseases such as Alzheimer's disease and Parkinson's syndrome has become a neuroscience Another research hotspot. [0003] The in vitro co-culture system that can be used to study the interaction between neurons and glial cells is a culture method established by Banker et al. in the 1990s for the purpose of culturing neurons. Co-cultivation method of metabolites ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/04C12N5/0793C12N5/079
CPCC12M35/08C12N5/0619C12N5/0622C12N2502/081C12N2502/086C12N2533/32
Inventor 陈振玲刘永锁王妍王伟于红燕
Owner 民航总医院
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