Mesenchymal stem cell chemical component limited serum-free culture medium and application thereof

A serum-free medium and mesenchymal stem cell technology, applied in the field of mesenchymal stem cell chemical composition-limited serum-free medium, can solve the problems of cell productivity, limited number of passages, infection of AIDS infectious diseases, and restrictions on the application of mesenchymal stem cells , to achieve the effect of eliminating instability, highly efficient differentiation potential, and maintaining differentiation potential

Inactive Publication Date: 2020-04-17
苏州依科赛生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional culture of adipose-derived mesenchymal stem cells requires the addition of fetal bovine serum. The cultured mesenchymal stem cells carry bovine serum albumin through endocytosis and uptake of bovine serum albumin, which can cause an immune response in the recipient. At the same time, bacteria, bacteria, The risk of viruses, and the limited yield and number of passages of cells cultured in serum greatly limit the application of mesenchymal stem cells
As well as the existing serum-free medium, human serum albumin, fibronectin and other components are added, affected by the source of raw materials, there is a risk of infection with AIDS, hepatitis B and other infectious diseases, which greatly improves the requirements for product quality control

Method used

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  • Mesenchymal stem cell chemical component limited serum-free culture medium and application thereof
  • Mesenchymal stem cell chemical component limited serum-free culture medium and application thereof
  • Mesenchymal stem cell chemical component limited serum-free culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The chemically defined serum-free medium for mesenchymal stem cells provided in this example consists of the following components: basal medium and additional components, the basal medium is DMEM / F12, and the additional components are recorded as MAX1 components, MAX1 components and The specific material composition of DMEM / F12 is as follows:

[0031] (1) The components contained in the MAX1 component and the content of each component are (based on the volume of the MAX1 component):

[0032]

[0033] (2) The components contained in the DMEM / F12 component (various components can be purchased from reagent companies such as Sigma and Aladdin) and the content of each component are (based on the volume of the DMEM / F12 component):

[0034]

[0035]

[0036]

[0037] In this example, the basal medium DMEM / F12 can also be purchased directly from the reagent company.

[0038] In this example, the basal medium DMEM / F12 was prepared by adding water and 1.2 g of sodium b...

Embodiment 2

[0041] This example provides verification of the effect of the serum-free medium defined by the chemical components of mesenchymal stem cells in Example 1.

[0042] In the experiments of this example, human adipose-derived mesenchymal stem cells were used. The cells were derived from the ATCC standard cell bank (PCS-500-011, Lot 80622175, ATCC), and the cells were used in all the following experiments.

[0043] 1. Experimental method

[0044] 1. Cell culture

[0045] Human adipose-derived mesenchymal stem cells, at 8000 / cm 2 The density was seeded on 100mm cell culture dishes, and the cell culture dishes used in the experiment were all Corning CellBinding surface treatment culture plates (Corning, 3335).

[0046] Add mesenchymal stem cell chemical composition-limited serum-free medium and control group containing bovine serum medium (DMEM / F12 medium (Sigma, M2906) with 10% fetal bovine serum (ExCellBio, FND500)) for continuous culture, the amount added About 15ml, every 48 ...

Embodiment 3

[0061] This example provides verification of the effects of various culture media. In the experiments of this example, human adipose-derived mesenchymal stem cells were used. The cells were derived from the ATCC standard cell bank (PCS-500-011, Lot 80622175, ATCC), and the cells were used in all the following experiments.

[0062] 1. Experimental method

[0063] 1. Preparation of culture medium:

[0064] Prepare following culture medium according to the technical method of embodiment 1

[0065] Numbering Volume ratio of medium solution (DMEM / F12 and MAX1 by volume ratio) M1 100:0.3 M2 100:0.5 M3 100:1 M4 100:2 M5 100:3 M6 100:4 M7 100:5

[0066] 2. Experimental methods and results

[0067] 1. Cell culture

[0068] Human adipose-derived mesenchymal stem cells, at 8000 / cm 2 The density was seeded on 100mm cell culture dishes, and the cell culture dishes used in the experiment were all Corning CellBinding surface treatment c...

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Abstract

The invention relates to a mesenchymal stem cell chemical component limited serum-free culture medium comprising a basic culture medium and additive components, and the additive components comprise: rhPDGF-BB, rhFGF-b, rhTGF-beta1, polyvinyl alcohol, alpha-ketoglutaric acid, lipopolysaccharide, recombinant fibronectin, recombinant human albumin, recombinant apo-transferrin, recombinant human insulin growth factors and hydrocortisone. The invention also provides application of the mesenchymal stem cell chemical component limited serum-free culture medium. The composition is clear and stable; instability of serum culture is eliminated, meanwhile, the factors have a synergistic effect, the adipose tissue-derived stromal cells can be efficiently amplified, the cell cycle is shortened, proliferation is rapid, the yield of the cells cultured for three generations reaches two times or above of that of serum culture, cell consistency is good, and the biological characteristics and differentiation potential of the cells can be efficiently maintained.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a serum-free medium with defined chemical components of mesenchymal stem cells and its application. Background technique [0002] Mesenchymal stem cells (MSCs) belong to the pluripotent stem cells of the mesoderm, mainly exist in connective tissue and organ mesenchyme, have the ability of self-renewal and multi-directional differentiation, and are widely found in umbilical cord tissue, fat, bone marrow, bone, etc. Muscle, periosteum, lung, liver, pancreas and other tissues as well as amniotic fluid and umbilical cord blood. Mesenchymal stem cells have a very strong ability to differentiate. Under specific conditions, they can proliferate and differentiate into various types of cells such as fat cells, bone cells, chondrocytes, nerve cells, and muscle cells. [0003] Among mesenchymal stem cells, adipose-derived mesenchymal stem cells (Human Umbilical Cord-Derived Mesenchyma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N2501/135C12N2501/115C12N2501/148C12N2501/15C12N2500/50C12N2500/30C12N2501/90C12N2500/25C12N2501/30
Inventor 于宝利李其雷陈旭陈刚孙芳
Owner 苏州依科赛生物科技股份有限公司
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