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Method for extracting anchorage-dependent cells and extracting device for method

An extraction method and a technology of adherent cells, applied in the field of biological cells, can solve problems such as separation and extraction difficulties, troublesome operation, and error-prone, and achieve the effects of reducing the possibility of error, convenient operation, and simple structure

Pending Publication Date: 2020-01-24
BONE MEDICAL TECH OF SUZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the separation and extraction of such cells is difficult and cumbersome. The current extraction methods either have many steps, are cumbersome to operate, and are prone to errors; The extraction method and the extraction device suitable for the method are of great significance for cell research

Method used

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  • Method for extracting anchorage-dependent cells and extracting device for method

Examples

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Effect test

example 1

[0032] A method for extracting adherent cells, comprising the following steps:

[0033] (1) Spread polyacrylamide hydrogel on the bottom surface of the cell culture well plate with a thickness of 1mm; use phosphate buffered saline (PBS buffer) to soak the polyacrylamide hydrogel for 7 days to eliminate the impact of the hydrogel on Cell toxicity.

[0034] (2) The polyacrylamide hydrogel treated in step (1) was irradiated with ultraviolet light for 20 minutes under the condition of soaking in PBS buffer solution, wherein: the wavelength of ultraviolet light is 313nm, and the ultraviolet irradiance is 356μW·cm -2 ; The hydrogel can be sterilized by ultraviolet irradiation, so that the cell culture can be carried out in a sterile environment;

[0035] After that, the PBS buffer was sucked away, and the polyacrylamide hydrogel was soaked with Sulfo-Sanpah solution for 1 hour. Sulfo-Sanpah solution is a cross-linking agent that can act as a bridge between polyacrylamide hydrogel a...

example 2

[0046] A method for extracting adherent cells, comprising the following steps:

[0047] (1) Spread polyacrylamide hydrogel on the bottom surface of the cell culture well plate with a thickness of 0.75mm; soak the polyacrylamide hydrogel in phosphate buffered saline (PBS buffer) for 15 days.

[0048] (2) The polyacrylamide hydrogel treated in step (1) was irradiated with ultraviolet light for 36 minutes under the condition of soaking in PBS buffer solution, wherein: the wavelength of ultraviolet light is 340nm, and the ultraviolet irradiance is 480μW·cm -2 ;

[0049] After that, the PBS buffer was sucked away, and the polyacrylamide hydrogel was soaked in Sulfo-Sanpah solution for 2.5 hours. - The concentration of Sanpah solution is 0.75mg / ml;

[0050] Afterwards, UV radiation for 40 minutes, and then let it stand for 2.5 hours;

[0051] After that, the Sulfo-Sanpah solution was sucked away, and the surface of the hydrogel was incubated with serum-free medium for 12 hours; ...

example 3

[0060] This example is basically the same as Example 2, except that the preparation steps of polyacrylamide hydrogel are slightly different, as follows:

[0061] a. acrylamide is configured into a 30wt% aqueous solution, methylenebisacrylamide is configured into a 3wt% aqueous solution, and ammonium persulfate is configured into a 5wt% aqueous solution for later use;

[0062] b. Weigh 2ml of acrylamide solution, 1.7ml of methylenebisacrylamide solution, 2.3ml of water, 120μl of ammonium persulfate solution and 6μl of tetramethylethylenediamine; mix well and add to Bio-Rad vertical glue pouring rack , standing at 30°C for 4 hours.

[0063] The obtained polyacrylamide hydrogel has an elastic modulus of 55.02kPa, has a certain degree of brittleness, is easy to separate and dig out, and has good light transmittance, which facilitates the observation of cells in the gel.

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Abstract

The invention relates to a method for extracting anchorage-dependent cells. The method comprises the following steps of preprocessing polyacrylamide hydrogel, culturing anchorage-dependent cells, integrally excavating the cells and the adhered gel, removing surface gel and the like. According to the method, an anchorage-dependent cell extracting device suitable for the extracting method is also arranged. The extracting method disclosed by the invention is convenient and easy to operate, and the extracting of the anchorage-dependent cells can be completed without tedious steps, so that the possibility of mistaking is reduced; besides, the adopted extracting device is simple in structure and low in cost; and in addition, the extracting method is small in damage to the anchorage-dependent cells, and has important significance to cell research.

Description

technical field [0001] The invention relates to the field of biological cells, in particular to a method for extracting adherent cells and an extraction device used in the method. Background technique [0002] Adherent cells, the growth of this type of cells must have a support surface that can be attached, and the cells can grow and reproduce on the surface by relying on the adhesion factors secreted by themselves or provided in the medium. Therefore, the separation and extraction of such cells are difficult and cumbersome. The current extraction methods either have many steps, are cumbersome to operate, and are prone to errors; The extraction method and the extraction device suitable for the method are of great significance for cell research. Contents of the invention [0003] The invention overcomes the deficiencies of the prior art, and provides a method for extracting adherent cells with simple operation and little cell damage and an extraction device suitable for th...

Claims

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Application Information

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IPC IPC(8): C12N5/00C12M3/00C12M1/34C12M1/26C12M1/00
CPCC12N5/0068C12M23/20C12M33/04C12M41/36C12M47/04C12N2533/30C12N2509/10
Inventor 罗宗平郭江博陈曦赛吉拉夫刘智慧王觉鸣
Owner BONE MEDICAL TECH OF SUZHOU CO LTD
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