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Method for large-scale production of fetal stem cell secretin

A stem cell and secretin technology, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve problems such as a large number of difficulties in culturing, promote adherence and clone formation, avoid differentiation, and reduce costs and needs. effect of space

Inactive Publication Date: 2019-02-01
姬松茸科研基金有限公司
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Although the market potential of mesenchymal stem cell secretin is great, there are great difficulties in its mass culture, mainly because stem cells require a large surface area for growth, and stem cells are very easy to differentiate, so we propose a large Method for large-scale production of secretin from fetal stem cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0021] Example: Culture of 400 million mesenchymal stem cells

[0022] First, the bioreactor is sterilized, and then the culture medium is injected into the bioreactor, and a certain amount of β-mercaptoethanol or monothioglycerol is added to the culture medium, and then 4 million mesenchymal stem cells are placed in the culture medium. Maintain the culture at a rotational speed of 40 to 50rmp, and add an appropriate amount of leukemia inhibitory factor. The leukemia inhibitory factor is added every 12 hours for about 8 days. When the number of embryonic stem cells reaches the standard, the embryonic stem cells are mixed with phosphate buffer and the microcarriers attached thereto were cleaned three times, then injected into the bioreactor with 100ml of culture fluid, continued to cultivate, collected the supernatant in the bioreactor after 12 hours, and obtained the protein concentrate by ultracentrifugation of the supernatant, and then Dilute the extracted protein concentrat...

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Abstract

The invention discloses a method for large-scale production of mesenchymal stem cell secretin, comprising the following steps: 1) stem cells are cultured through a culture solution in a bioreactor, and the internal conditions of the bioreactor are 37 DEG C and 5 % carbon dioxide gas; 2) the stem cells obtained in the Step 1 are washed with a phosphate buffer in the bioreactor; then a cell culturemedium buffer is injected into the bioreactor to continuously culture the stem cells for 12 hours; and 3) a supernatant in the bioreactor is collected, and the supernatant undergoes ultracentrifugation to obtain a protein concentrate. According to the method for large-scale production of mesenchymal stem cell secretin, enzyme in cells and sulfhydryl groups in protein can be protected from being oxidized by adding beta-mercaptoethanol or monothioglycerol. With the addition of the leukemia inhibitory factor, the leukemia inhibitory factor can avoid stem cell differentiation and maintain stem cell pluripotency.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for large-scale production of fetal stem cell secretin. Background technique [0002] Stem cells are a type of pluripotent cells with self-replicating ability. Under certain conditions, it can differentiate into a variety of functional cells. Stem cells are divided into embryonic stem cells and adult stem cells according to their developmental stage. Stem cells are divided into three categories according to their developmental potential: totipotent stem cells, pluripotent stem cells, and unipotent stem cells (multipotent stem cells). Stem cells are not fully differentiated and immature cells that have the potential to regenerate various tissues and organs and the human body. They are called "universal cells" in the medical field, and stem cells are the cells of origin. Stem cells are cells with proliferation and differentiation potential, have the ability of self-renewal ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00C12N5/0775
CPCC12P21/00C12N5/0662C12N2500/44C12N2501/235
Inventor 李刚王彬林思恩李郁伟
Owner 姬松茸科研基金有限公司
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