Adipose-derived stem cell preparation and preparation method thereof

An adipose stem cell and preparation technology, which is applied in pharmaceutical formulations, medical preparations with non-active ingredients, and medical preparations containing active ingredients, etc. problems, to achieve the effect of improving survival, good survival and viability, maintaining viability and viability

Inactive Publication Date: 2015-07-15
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, adipose stem cells are very fragile, and it is difficult to ensure the survival rate when they are made into preparations, and if the survival rate of adipose stem cells in the preparation is insufficient, it will not be able to exert a good drug effect

Method used

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  • Adipose-derived stem cell preparation and preparation method thereof
  • Adipose-derived stem cell preparation and preparation method thereof
  • Adipose-derived stem cell preparation and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Preparation of Adipose Stem Cells

[0048] 1) Separation

[0049] Body adipose tissue was divided into 50mL centrifuge tubes, 20mL in each tube. Add an equal volume of 0.5% type Ⅰ collagenase (final concentration: 0.25%) to each tube, mix thoroughly, seal, transfer to a constant temperature air shaker, and digest for 1 hour at 37°C and 100R. Add 4mL of FBS to each tube to stop digestion and mix well. Centrifuge at 1500rpm / min for 5min. Discard the upper two layers of liquid, add 40mL PBS to each tube to resuspend, and resuspend the cells. Centrifuge at 1500rpm / min for 5min, discard the supernatant to obtain fat stem cells, which are seeds.

[0050] 2) Cultivate

[0051] Add DMEM-F12 medium containing 15% FBS to resuspend the cells at 1×10 5 individual / mL inoculation. Gently shake the culture dish back and forth to distribute the cell suspension evenly at the bottom of the flask; transfer to 37°C, 5% CO 2 , cultured in an incubator with a saturated humi...

Embodiment 2～4

[0058] Albumin and low-molecular-weight hyaluronic acid were added to physiological saline to obtain a preparation matrix, and the washed adipose stem cells prepared in Example 1 were resuspended in the matrix. The amounts of albumin, hyaluronic acid, and adipose stem cells in each embodiment are shown in Table 1:

[0059] Table 1 Examples 2-4

[0060]

Embodiment 5

[0069] Embodiment 5 preparation quality evaluation

[0070] Quality identification was carried out on the stem cell preparations prepared in Examples 2-4 and Comparative Examples 1-6. Each sample was checked 3 times. The method is:

[0071] After the preparation is made, the freshly prepared preparation is tested within 3 hours, and the detection content includes: stem cell morphology, survival rate, and the content of bacteria in the preparation. Among them, trypan blue staining was used for cell morphology detection and cell viability detection. The test results are shown in Table 4. image 3 The result of trypan blue staining within 3 hours of preparation of the adipose stem cell preparation provided in Example 4 of the present invention is shown; in the adipose stem cell preparation prepared in other embodiments, the morphology of the stem cells is similar to this.

[0072] Table 4 Test results within 3 hours after preparation

[0073]

[0074] Note: A means that t...

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Abstract

The invention relates to the field of biology and medicines, and especially relates to an adipose-derived stem cell preparation and a preparation method thereof. The adipose-derived stem cell preparation comprises adipose-derived stem cells, hyaluronic acid, alhumin and normal saline. The adipose-derived stem cells can maintain good survival rate and activity, and do not differentiate under the protection of hyaluronic acid, alhumin and normal saline in the adipose-derived stem cell preparation. The preparation method of the adipose-derived stem cell preparation provided by the invention can avoid damages of the adipose-derived stem cells, and improves the survival rate of the adipose-derived stem cells in the preparation. Experiments show that the adipose-derived stem cell preparation can maintain the survival rate of the adipose-derived stem cells above 99%, and allows the cells to have uniform size and shuttle-shaped form. After the adipose-derived stem cells are preserved at 0-4DEG C for 72h, the cells still maintain good form and have no volume enlargement or other abnormal phenomena, the survival rate still maintains above 95%, and the adipose-derived stem cells still maintain the characteristics of stem cells, and do not differentiate.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to an adipose stem cell preparation and a preparation method thereof. Background technique [0002] Osteoarthritis (OA) is a chronic and disabling disease caused by the interaction of mechanical and biological factors, causing lesions in articular chondrocytes and extracellular matrix; it mostly occurs in middle-aged and elderly people, and the prevalence rate increases significantly with age high. Current treatment methods include surgery and drug intervention, but it is still difficult for patients to restore normal cartilage function. Therefore, the repair of cartilage damage has always been a thorny issue in clinical research. In recent years, bone marrow mesenchymal stem cells have been used as repair cells for osteoarthritis. However, in order to obtain bone marrow stem cells, the bone marrow must be extracted through puncture, and then separated, purified and extracted. In addi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K47/42A61K47/36A61P19/02
Inventor 王一飞陈海佳葛啸虎卢瑞珊王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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