Human adipose tissue-derived stromal cell frozen stock solution

A technology of high-quality stem cells and cryopreservation solution, which is applied in the field of stem cells, can solve the problems of not being suitable for clinical application, introducing polluting allergens, and affecting treatment results, so as to avoid the risk of pollution and allergens, high clinical safety, and maintain stem cells. The effect of the characteristic

Inactive Publication Date: 2015-04-01
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, fetal bovine serum is a heterogeneous substance with complex components and the risk of introducing contamination and allergens, so it is not suitable for clinical application
Especially in cell therapy, the presence of heterologous proteins may cause unnecessary or even unknown adverse reactions, seriously affecting the treatment results

Method used

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  • Human adipose tissue-derived stromal cell frozen stock solution
  • Human adipose tissue-derived stromal cell frozen stock solution
  • Human adipose tissue-derived stromal cell frozen stock solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Primary isolation culture and subculture of human adipose-derived mesenchymal stem cells

[0027] The adipose tissue obtained by liposuction was washed with PBS until clarified, and an equal volume of preheated 0.5% type I collagenase was added to the washed adipose tissue. The mixture was transferred to a constant temperature air shaker and digested at 37°C and 100rpm for 50min. After the adipose tissue block was viscous and fine granular, it was centrifuged at 1500r / min for 5min. Carefully remove the upper miscellaneous liquid, leave 5mL suspension at the bottom, add an appropriate amount of PBS to dilute the effect of collagenase. Centrifuge again at 1500r / min for 5min to collect the cells. Resuspend cells with complete medium at 1×10 5 Inoculate 10 cm culture dishes at a density of cells / mL, and add 100 μL of 1 μg / mL epithelial growth factor (EGF) to each dish. Move cells to CO 2 In the incubator, change the medium after culturing for 48 hours to rem...

Embodiment 2

[0028] Embodiment 2, the preparation of autologous plasma

[0029] Extract 50mL of human autologous venous blood into an anticoagulant blood collection tube, transfer it to a 50mL centrifuge tube, centrifuge horizontally at 2000r / min for 15min, and absorb the supernatant for later use, which is autologous plasma.

Embodiment 3

[0030] Embodiment 3, preparation of cell cryopreservation solution

[0031] According to the formula shown in Table 1, the cell cryopreservation solution for cryopreservation cells was prepared. in

[0032]

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PUM

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Abstract

The invention relates to the field of stem cells, discloses a frozen stock solution of stem cells, and in particular discloses a human adipose tissue-derived stromal cell frozen stock solution which consists of human plasma and dimaethyl sulfoxide. Compared with an ordinary cell frozen stock solution, the human adipose tissue-derived stromal cell frozen stock solution disclosed by the invention is relatively high in clinical security as no non-human source serum or protein is provided and the risk that contamination or allergen is introduced is avoided. The experience shows that compared with the ordinary cell frozen stock solution, the human adipose tissue-derived stromal cell frozen stock solution disclosed by the invention is relatively high in motility rate of cells in frozen stock, good stem cell characteristics are maintained, and human fat stem cells can be preserved and applied for a long time.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a cryopreservation solution for stem cells, especially a cryopreservation solution for human adipose-derived mesenchymal stem cells. Background technique [0002] Stem cells are a type of pluripotent cells with self-renewing ability. Under certain conditions, it can differentiate into a variety of functional cells, which are called "universal cells" in the medical field. Stem cells are divided into embryonic stem cells (ES cells) and adult stem cells (somatic stem cells) according to their developmental stages. Stem cells are divided into three categories according to their developmental potential: totipotent stem cells (TSCs), pluripotent stem cells (pluripotent stem cells) and unipotent stem cells (unipotent stem cells). [0003] Adipose Mesenchymal Stem Cells (ADSC) are adult pluripotent stem cells that exist in adipose tissue, and can differentiate into various tissue cells such a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 王一飞陈海佳葛啸虎麦锦连王小燕马岩岩
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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