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Method for culturing mesenchymal stem cell secretin

A technology of mesenchymal stem cells and secretin, applied in the field of preparation of mesenchymal stem cell secretin, can solve the problems of protein loss, high cost, high cost, etc., and achieve the effect of increasing production, high compatibility, improving safety and effect

Active Publication Date: 2014-05-21
ASIA PACIFIC STEM CELL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. Cell culture flasks are mainly used to collect secretin. Due to the attachment growth characteristics of mesenchymal stem cells, the number of cells required for mass production of secretin will take a lot of culture flasks and space, and it will take time and cost to cultivate. high;
[0006] 2. When collecting secretin, a large amount of culture medium is required, and the cost is very high;
[0007] 3. The protein in secretin will be lost in a large amount when it is concentrated, and the product contains impurities such as high salt and cell culture fluid

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: cultivate 200 million adipose-derived mesenchymal stem cell secretin

[0029] First, 4 million adipose-derived mesenchymal stem cells were cultured in T-150 culture flasks containing cell culture medium, which contained 10% bovine serum, 1% antibiotics, and alpha-MEM medium. 30 million adipose-derived mesenchymal stem cells were produced; then the above-mentioned adipose-derived mesenchymal stem cells were transferred to a 3-liter bioreactor with 3 grams of microcarriers and 400 milliliters of cell culture medium, maintained at a speed of 40 rpm Cultivation: Inject 200 ml and 500 ml of cell culture solution into the bioreactor at the 24th hour and the 72nd hour respectively. Continue cell culture until the number of adipose-derived mesenchymal stem cells reaches 200 million (about 6 days); When the number of adipose-derived mesenchymal stem cells reached the standard, the adipose-derived mesenchymal stem cells and their attached microcarriers were washed t...

Embodiment 2

[0030] Embodiment 2: cultivate 200 million dental pulp mesenchymal stem cell secretin

[0031] The preparation method of this example is roughly the same as that of Example 1, except that dental pulp mesenchymal stem cells are used instead of adipose-derived mesenchymal stem cells, and the growth time in a 3-liter bioreactor can be reduced from 9 days to 6 days in total, while The time for collecting secretin from dental pulp mesenchymal stem cells can be increased to 72 hours.

Embodiment 3

[0032] Embodiment 3: cultivate 20,000,000 umbilical cord mesenchymal stem cells secretin

[0033] The preparation method of this example is roughly the same as that of Example 1, except that umbilical cord mesenchymal stem cells are used instead of adipose-derived mesenchymal stem cells, and 4 million freshly thawed umbilical cord mesenchymal stem cells are directly placed in a microcarrier containing 0.8 g and 300 ml of culture medium in a 500 ml bioreactor, and the cell culture medium was renewed in the bioreactor for 72 hours, and the growth time in the bioreactor could be increased from 9 days to 10 to 12 days in total.

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Abstract

The invention provides a method for preparing mesenchymal stem cell secretin with low production cost and high product purity, a bioreactor technology is used for replacing a traditional cell culture bottle to prepare mesenchymal stem cell secretin, the cost and the required space can be greatly reduced; the used mesenchymal stem cells enables high compatibility, the mesenchymal stem cells is continuously stimulated in a safe mode to secrete the protein, and the output of mesenchymal stem cell secretin can be enhanced.

Description

technical field [0001] The invention relates to the preparation technology of mesenchymal stem cell secretin. Background technique [0002] Existing mesenchymal stem cell secretin beauty products (such as apple stem cells) have been accepted by the public, and the academic circles are conducting in-depth discussions on the potential medical use of secretin from mesenchymal stem cells. In recent years, secretin has been The literature of positive medical effects emerged, such as improvement in skin wrinkles, heart failure, brain degeneration, eyesight, life extension, etc. In recent years, a large number of documents have also pointed out the other potential medical value of secretin. The most in-depth study is its function of improving heart failure. Studies have pointed out that exosomes contained in stem cell secretions have certain medical applications such as Disclosed in US Patent 2011 / 0003008A1. Although the market potential of secretin from mesenchymal stem cells is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/00
Inventor 周向荣邓铭权陈詠敏
Owner ASIA PACIFIC STEM CELL SCI
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