Induced differentiation method of 3T3-L1 preadipocytes line

A 3T3-L1, 1.3T3-L1 technology, applied in the field of induction and differentiation of 3T3-L1 preadipocyte line, can solve the problems of difficult to provide quantity for researchers, low conversion rate of adipocytes, long induction period, etc., to shorten the differentiation process, prolonged action time, stable conversion rate

Inactive Publication Date: 2015-11-25
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the deepening of research, it is found that the traditional "cocktail" induction method has disadvantages such as long induction period, low and uneven fat cell conversion rate, etc.
In addition, the induction method will also be affected by the passage number of cells and the type of cell culture dish
Therefore, it is difficult to provide researchers with adipocytes with consistent numbers and stable differentiation

Method used

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  • Induced differentiation method of 3T3-L1 preadipocytes line
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  • Induced differentiation method of 3T3-L1 preadipocytes line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Recovery of 3T3-L1 preadipocyte line: remove the cell line from liquid nitrogen, immediately place it in a 37±2°C water bath, take it out when the cell fluid is completely dissolved, centrifuge at 1000 rpm for 5 min at room temperature, discard the supernatant, and add to the culture The resuspended cells and culture medium were sucked into the culture dish, and the shape and number of cells were observed under the microscope, so that the cell suspension was 3×10 5 cells / ml into a culture vessel, which was then placed in 5% ± 1% CO 2 cultured in a constant temperature incubator at 37±2°C; wherein, the culture medium is a high-glucose DMEM medium containing 10% calf serum and 1% penicillin-streptomycin;

[0042]Cultivation of 3T3-L1 preadipocyte line: cells adhered under the microscope, shuttled and translucent, the cells were replaced with the culture medium every 3 days, the culture medium was tilted, the culture medium was removed with a pipette, and added along the w...

Embodiment 2

[0060] Recovery of 3T3-L1 preadipocyte line: remove the cell line from liquid nitrogen, immediately place it in a 37±2°C water bath, take it out when the cell fluid is completely dissolved, centrifuge at 1200 rpm for 3 min at room temperature, discard the supernatant, and add to the culture The resuspended cells and culture medium were sucked into the culture dish, and the cell morphology and number were observed under the microscope, so that the cell suspension was 6×10 5 cells / ml into a culture vessel, which was then placed in 5% ± 1% CO 2 cultured in a constant temperature incubator at 37±2°C; wherein, the culture medium is high-glucose DMEM medium containing 10% calf serum and 1.1% penicillin-streptomycin;

[0061] Cultivation of 3T3-L1 preadipocyte line: the cells adhered to the wall under the microscope, were shuttled and translucent, the cells were replaced with the culture medium every 2 days, the culture medium was tilted, the culture medium was removed with a pipette...

Embodiment 3

[0072] Recovery of 3T3-L1 preadipocyte line: remove the cell line from liquid nitrogen, immediately place it in a 37±2°C water bath, take it out when the cell fluid is completely dissolved, centrifuge at 1000 rpm for 5 min at room temperature, discard the supernatant, and add to the culture The resuspended cells and culture medium were sucked into the culture dish, and the shape and number of cells were observed under the microscope, and the cell suspension was 1×10 5 cells / ml into a culture vessel, which was then placed in 5% ± 1% CO 2 Cultured in a constant temperature incubator at 37±2°C; wherein, the culture medium is high-glucose DMEM medium containing 10% calf serum and 0.9% penicillin-streptomycin;

[0073] Cultivation of 3T3-L1 preadipocyte line: cells adhered under the microscope, shuttled and translucent, the cells were replaced with the culture medium every 3 days, the culture medium was tilted, the culture medium was removed with a pipette, and added along the wall...

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Abstract

The invention relates to the field of the induction of adipocytes, in particular relates to an induced differentiation method of 3T3-L1, and the method comprises the steps of performing resuscitation, culturing, passage on the 3T3-L1 preadipocytes line until culture mediums is overgrown with preadipocytes, and after continuously culturing for 36 to 48 hours, adding a high glucose DMEM culture solution containing an inducer and 10 percent of fetal calf serum to carry out the induced differentiation for 72 to 96 hours; the inducer is prepared from 0.9 to 1.1 mu. M of dexamethasone, 1.0mM of IBMX, and 1.8 to 2.0 mu. M of insulin; adding the high glucose DMEM culture solution containing the insulin and the fetal calf serum to induce continuously for 48 to 96 hours; and then, changing the culture solution once every 48 hours, and mature adipocytes can be used from the 8th to 10th day after the inducer is added. The induced differentiation method can shorten the whole induced process of the 3T3-L1 preadipocytes line, the conversion ratio of the adipocytes is stable, and is not influenced by the number of passages and the types of cell culture dishes, and the error of the conservation rate is within 5%.

Description

technical field [0001] The present invention relates to the field of adipocyte induction, in particular, to a method for inducing differentiation of 3T3-L1 preadipocyte line. Background technique [0002] With the improvement of people's living standards, the incidence of obesity and related diseases is increasing, and the resulting family and socioeconomic burdens are increasing. Metabolism and dysfunction of adipocytes is one of the core mechanisms leading to obesity, and research on adipocytes has become a current research hotspot. In in vitro studies, adipocyte models are important tools for studying fat metabolism and its function. [0003] The 3T3-L1 preadipocyte line is derived from the fibroblast population of mouse placenta. Studies have found that it is a precursor cell that can proliferate and differentiate into adipocytes under certain conditions. At present, it has become the most commonly used cell line to induce differentiation into adipocytes. [0004] Fo...

Claims

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Application Information

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IPC IPC(8): C12N5/077
Inventor 刘新农张锦航李天佳王占启刘昌伟
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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