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Adipose cell large-scale culture method and device

A culturing method and a fat cell technology are applied in the field of a large-scale culturing method for fat cells and a culturing device thereof, and can solve the problems of harsh operating conditions, limited application of microcarriers, and high requirements

Inactive Publication Date: 2019-08-13
张永国
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The method for culturing cells with microcarriers in the prior art has complex processes, harsh operating conditions, low cell survival rate, low inoculation density, and high requirements for the microcarrier itself. It is necessary to use special microcarriers to ensure normal cell culture. Indirectly increase the operating cost and limit the application of microcarriers

Method used

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  • Adipose cell large-scale culture method and device
  • Adipose cell large-scale culture method and device
  • Adipose cell large-scale culture method and device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] The large-scale culture method of fat cells is carried out according to the following steps:

[0071] 1) After digesting the adherent adipocytes, inoculate them in a biological culture device 20 and co-culture with GE microcarriers. The inoculation density of adipocytes is 8*10 3 cell / ml, the final concentration after inoculation is 0.2wt% (the total concentration of GE microcarrier and cells to be cultured);

[0072] 2) culture fluid is housed in the sterile culture bag 10, and a joint 50 is installed on the side wall of the sterile culture bag, which is connected with the inlet of the biological incubator 20 through the joint, so as to transport nutrients through the adjustable variable speed pump 30, and the biological culture The outlet of the device 20 is provided with a joint 50, and the culture solution in the biological incubator 20 is discharged through the variable speed pump 30;

[0073] 3) After stirring and culturing at a slow speed of 10-15rpm for 24 hour...

Embodiment 2

[0076] The large-scale culture method of fat cells is carried out according to the following steps:

[0077] 1) Treat the microcarriers: prepare 4L of PBS 0.01M pH7.4, autoclave at 121°C for 15min, soak the Cytodex 1GE microcarriers with the above PBS overnight, wash 3 times, the beads settle to the bottom, discard the broken sterilized supernatant After that, the medium was used to equilibrate overnight at room temperature, and the medium was changed once in the middle. Aliquot 10ml / tube, store at 4°C, take 1ml and add 100ml medium when used;

[0078] 2) After digesting the adherent adipocytes, inoculate them in a biological culture device 20 to co-culture with microcarriers, and the inoculation density of adipocytes is 8.5*10 3 cell / ml, the final concentration after inoculation is 0.8wt% (total concentration of GE microcarrier and cells to be cultured);

[0079] 3) culture fluid is housed in the sterile culture bag 10, and the side wall of the sterile culture bag is equipp...

Embodiment 3

[0083] The large-scale culture method of fat cells is carried out according to the following steps:

[0084]1) Treat the microcarriers: prepare 4L of PBS 0.01M pH7.4, autoclave at 121°C for 15min, soak the Cytodex 1GE microcarriers with the above PBS overnight, wash 3 times, the beads settle to the bottom, discard the broken sterilized supernatant After that, the medium was used to equilibrate overnight at room temperature, and the medium was changed once in the middle. Aliquot 10ml / tube, store at 4°C, take 1ml and add 100ml medium when used;

[0085] 2) After digesting the adherent adipocytes, inoculate them in a biological culture device 20 to co-culture with microcarriers, and the inoculation density of the adipocytes is 8.4*10 3 cell / ml, the final concentration after inoculation is 0.1wt% (the total concentration of GE microcarrier and cells to be cultured);

[0086] 3) culture fluid is housed in the sterile culture bag 10, and the side wall of the sterile culture bag is...

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Abstract

The invention provides an adipose cell large-scale culture method and device. The culture method includes the following steps that to-be-cultured cells and a sterilized GE micro-carrier are inoculatedinto a serum-containing culture medium and subjected to stirring culture for 24 hours or longer, wherein the total concentration of the inoculated GE micro-carriers and the to-be-cultured cells is 0.1-1wt%, and the inoculation density of the to-be-cultured cells is (8-8.5)*10<3> cell / ml. The culture method is simple to operate, operation conditions are relatively mild, the cell survival rate is high, a reference method for subsequent large-scale culture of adipose cells is provided, and the method is worthy of wide application and popularization.

Description

technical field [0001] The invention relates to the field of adipocyte culture, in particular to a large-scale culture method of adipocytes and a culture device thereof. Background technique [0002] Every adult body contains about 30 billion white fats, whose function is to store energy in the form of fat cells. Each fat cell contains triglycerides, commonly known as fat globules. When the amount of fat globules becomes larger, the volume of fat cells will expand, resulting in obesity; on the contrary, when triglycerides are burned, the cells will shrink and the body will lose weight. Under normal circumstances, the number of fat cells does not increase after puberty. The distribution of body fat is partly determined by genetic and hormonal influences. For example, women's subcutaneous fat mostly accumulates in the lower abdomen, buttocks and thighs, while men accumulate it in the upper abdomen and waist. [0003] Microcarriers refer to microbeads with a diameter of 60-2...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12M3/02C12M1/24
CPCC12N5/0653C12M23/14C12M23/08C12M23/58C12M29/00C12N2531/00
Inventor 张永国何君
Owner 张永国
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