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Evaluation system of anti-aging effect of autologous adipose-derived stem cell

A fat stem cell, anti-aging technology, applied in the field of biotechnology and regenerative medicine, can solve the problems that hinder the anti-aging of autologous fat stem cells

Active Publication Date: 2012-06-06
CELLULAR BIOMEDICINE GRP SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Therefore, despite the rapid development of adipose stem cells for anti-aging, the establishment of an evaluation system for autologous adipose stem cells is hindered due to the effects of the internal and external environment of the organism. Anti-aging evaluation method and system of fat stem cells

Method used

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  • Evaluation system of anti-aging effect of autologous adipose-derived stem cell
  • Evaluation system of anti-aging effect of autologous adipose-derived stem cell
  • Evaluation system of anti-aging effect of autologous adipose-derived stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0146] Obtaining of stromal vascular fraction (SVF)

[0147] 1. Obtaining Adipose Tissue

[0148] Wipe the outer wall of the container containing adipose tissue with 75% alcohol, draw the adipose tissue from the human body with a sterile needle, and move it to the receiving container within 1 hour.

[0149] 2. Aliquot Adipose Tissue

[0150] Each culture bottle is divided into 50ml of adipose tissue, use a 10ml pipette to absorb the lower layer of red liquid in the fat collection bottle, discard it, and then divide the remaining upper layer of fat after mixing.

[0151] 3. Wash adipose tissue to remove blood cells

[0152] Add 100ml of sodium chloride injection to the culture bottle, tighten the cap, shake vigorously for 3 minutes to fully wash the fat tissue, then stand still for 3-5 minutes to separate the different phases, and suck off the lower aqueous phase; repeat the above operation three times until the lower layer The liquid is clearer.

[0153] 4. Collagenase I D...

Embodiment 2

[0160] Adipose stem cell planting and culture

[0161] 1. Cell Seeding

[0162] Add 20ml of medium to the centrifuged SVF, and mix well. According to the area of ​​the culture bottle, use the tissue block method to plant cells, and inoculate according to the amount of fat obtained by inoculating 0.16ml of liposuction per square centimeter. For every 100ml of adipose tissue, Ultimately 8 T75 flasks can be inoculated.

[0163] 2. Primary Cell Culture

[0164] Place the flat culture bottle in a carbon dioxide constant temperature and humidity incubator for cultivation, the cultivation condition is 37±0.5°C, and the volume fraction of carbon dioxide is 5±0.2%. In the 24th hour of the primary culture, a full volume of medium was changed. After that, the medium was changed every 3 days, and the culture was carried out in a constant temperature and humidity incubator with carbon dioxide.

[0165] 3. Primary Cell Harvesting

[0166] After about 7 days, the area percentage of the ...

Embodiment 3

[0171] Antigen marker detection

[0172] SVF and cultured adipose stem cells were collected into centrifuge tubes, and the cell suspension was adjusted to a density of 1×10 5 mL -1 , 800rpm (120g), centrifuge for 5min, discard the supernatant, wash and resuspend the cells with cold D-Hanks at 4°C, and centrifuge the cell suspension again at 800rpm for 5min, then discard the supernatant. Then the cells were resuspended to 1 mL with D-Hanks, 5 μl of antibody was added, protected from light, and placed on ice for 30 min. Rinse with D-Hanks, centrifuge, discard the supernatant, and repeat the washing process twice to ensure that the unbound antibody is removed. Finally, add about 200 μl of D-Hanks to make a suspension, and detect it with a flow cytometer.

[0173] The added antibodies were: human anti-CD29, CD73, CD49d, CD90, CD14, CD34, CD45, CD34, Actin and HLA-DR. Antigen detection results are shown in Table 1.

[0174] Table 1

[0175]

[0176] It shows that the analy...

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Abstract

The invention provides an evaluation system of anti-aging effect of autologous adipose-derived stem cell. The system comprises that serology detection is carried out on an individual re-transfused with adipose-derived stem cells a period after a re-transfusion; and the serology detection comprises one or more indexes selected from the group of vitamin D, high density lipoprotein, low density lipoprotein, hormothyrin, testosterone and estradiol. The evaluation system of the invention can objectively evaluate anti-aging effect of adipose derived stem cell and has advantages of accuracy, rapidness and thrift.

Description

technical field [0001] The invention relates to the fields of biotechnology and regenerative medicine. Specifically, the present invention relates to an evaluation system for the anti-aging effect of autologous adipose stem cells. Using the system of the present invention, the anti-aging effect of adipose stem cells can be evaluated accurately, objectively and in a timely manner. Background technique [0002] In the whole life process of human beings from embryonic development, life birth to maturity, aging to death, tissue and organ damage and functional decline will inevitably occur. In the normal physiological process, a large number of cells die in the human body every day, accompanied by the regeneration of a large number of cells, in order to maintain the integrity of various tissues and organs and ensure their functions are in a normal state. If the balance between cell death and regeneration is out of balance, the increase of dead cells or the decrease of new cells ...

Claims

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Application Information

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IPC IPC(8): G01N33/96G01N33/53A61K35/28
CPCG01N33/92A61K35/28A61K48/00G01N33/743G01N33/82G01N33/96A61K35/12
Inventor 曹卫吕伟麟张丽张露亿张炳强
Owner CELLULAR BIOMEDICINE GRP SHANGHAI
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