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Two-time shifting cell migration assay method

An assay method and cell migration technology, applied in the field of cell biology, can solve problems such as high cost, unclean erasing, and time-consuming, and achieve the effects of avoiding errors, shortening cell migration distance, and eliminating manual delimitation.

Active Publication Date: 2021-02-09
PUTIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the need to manually erase the cells retained in the cup, it takes a long time and cannot avoid the problem of unclean erasure. At the same time, the price of the porous polycarbonate membrane cup is relatively expensive, and the number of porous polycarbonate membranes required for the experiment is large, and there is a cost. too high problem
The above two methods usually need to be cultured for 2-3 days during the measurement of migration, and the influence of cell proliferation cannot be avoided

Method used

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  • Two-time shifting cell migration assay method
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  • Two-time shifting cell migration assay method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] refer to Figure 1-2 , twice shift cell migration assay method specifically comprises the following steps:

[0032] Step 1. Place the nipple slides in the culture wells of the first 96-well cell culture plate in turn, and then insert 100 μL of cells to be tested in each well (culture well of the culture plate) for cell culture, and form 2-4x10 cells the next day. 4 density of cells / well;

[0033] Step 2. After the cell culture is over (80-100% full), take out the papilla slides one by one, pick up the papilla slides with sterilized tweezers and place them in the culture wells of the second 96-well cell culture plate one by one, completing the second step. Translocate once, then add cell culture medium and test substances (such as organic reagents, inorganic reagents, peptides, biologically active substance inhibitors, stimulants, etc.), and cultivate for 12-24 hours. At this time, the cells migrate, and the migrated cells pass through the nipple The edge of the slide ...

Embodiment 2

[0039] refer to Figure 3-4 , twice shift cell migration assay method specifically comprises the following steps:

[0040]Step 1. Place nipple slides in the culture wells of the first 48-well cell culture dish, and then insert 200 μL of cells to be tested into each well (culture wells of the culture dish) for cell culture, and form 6-8x10 cells the next day. 4 density of cells / well;

[0041] Step 2. After the cell culture is over, take out the papilla slides one by one, place 48 papilla slides in the culture wells of the second 48-well cell culture dish in turn, complete the first shift, and then add the cell culture medium and the test substance (such as organic reagents, inorganic reagents, peptides, biologically active substance inhibitors, stimulants, etc.), cultured for 12-24 hours, at this time the cells migrated;

[0042] Step 3. Take the papilla slides out of the second 48-well cell culture dish, and complete the second shift. Those left in the second cell culture di...

Embodiment 3

[0047] The difference between embodiment 3 and embodiment 1 is:

[0048] Both the first cell culture dish and the second cell culture dish are 24-hole cell culture dishes;

[0049] The material of the nipple slide is polyethylene resin;

[0050] The through hole is strip-shaped, the thickness of the papilla slide is 0.5 mm, the papilla slide is cuboid, and the length of the papilla slide is less than the diameter of the culture hole of the 24-hole cell culture dish.

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Abstract

The invention belongs to the technical field of cell biology, and discloses a two-time shifting cell migration assay method. The method comprises the following steps: placing a nipple slide in a culture hole of a first cell culture disc, and inoculating a cell to be determined; taking out the nipple slide after culturing, and placing the nipple slide in a culture hole of a second cell culture disc; adding a cell culture solution and an experimental substance; and taking out the nipple slide, and performing quantification through an XTT colorimetric method or an MTT colorimetric method. The nipple slide comprises a slide body and a mastoid arranged at the center of the slide body, and a plurality of through holes are formed in the slide body; and the nipple slide is made of polymer resin. The two-time shifting cell migration assay method is slightly influenced by manual operation, the cell migration distance is short, the time is short, the method is not easily influenced by cell proliferation, used equipment is generally arranged in laboratories, the required cost is low, and common laboratories can autonomously design and finish the required cell migration assay according to the provided two-time shifting cell migration assay method.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and in particular relates to a method for measuring the migration of twice-shifted cells. Background technique [0002] Cell migration assays are one of the fundamental methods for probing the behavioral capabilities of cells in cell biology research. It is widely used in cell biology research. Through the study of cell migration ability, we can understand the mechanism of physiological and pathological processes, such as angiogenesis, embryonic development, immune cell function, tumor cell migration, wound healing, etc. [0003] There are two methods commonly used to measure cell migration. The first is the wound healing experiment. In this method, the cells planted on the glass slide are artificially scraped off a 1-2mm wide blank area, and the boundary is marked with a marker pen, and the slide is placed in the culture medium of the test condition. Cultivate for a certain period of tim...

Claims

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Application Information

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IPC IPC(8): C12Q1/06G01N21/31
CPCG01N33/5029G01N21/31
Inventor 朱棠
Owner PUTIAN UNIV
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