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Mesenchymal stem cell serum-free culture medium and application thereof

A serum-free medium and mesenchymal stem cell technology, applied in the field of mesenchymal stem cell serum-free medium, can solve the problems of cell productivity, limited number of passages, and limit the application of mesenchymal stem cells, so as to eliminate instability and maintain Biological characteristics and differentiation potential, the effect of high-efficiency amplification

Active Publication Date: 2020-03-31
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

Traditional mesenchymal stem cell culture 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 immune responses in the recipient. At the same time, bacteria and viruses carried by heterologous serum can be introduced. In addition, the cell yield and passage times of serum culture are limited, which greatly limits the application of mesenchymal stem cells

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The 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 DMEM / F12 The specific material composition is as follows:

[0029] (1) The components contained in the MAX1 component and the content of each component are (based on the volume of the MAX1 component):

[0030]

[0031]

[0032] (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):

[0033]

[0034]

[0035] In this example, the basal medium DMEM / F12 can also be purchased directly from the reagent company.

[0036]In this example, the basal medium DMEM / F12 was prepared by adding water and 1.2 g of sodium bicarbonate (Sigma, S5...

Embodiment 2

[0039] This example provides verification of the effect of the serum-free medium for mesenchymal stem cells in Example 1.

[0040] In the experiments of this example, human umbilical cord mesenchymal stem cells were used. The cells were derived from the ATCC standard cell bank (PCS-500-010, Lot 63739206, ATCC), and the cells were used in all the following experiments.

[0041] 1. Experimental method

[0042] 1. Cell culture

[0043] Human umbilical cord 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 pre-coated and contained the following human adhesion molecules: recombinant human fibronectin (rhFibronectin).

[0044] Add mesenchymal stem cell serum-free medium and control medium containing bovine serum (DMEM / F12 medium (Sigma, M2906) with 10% fetal bovine serum (ExCellBio, FND500)) for continuous culture, the amount added is about 15ml , per 48 ～ The culture medium was c...

Embodiment 3

[0059] This example provides verification of the effects of various culture media. In the experiments of this example, human umbilical cord mesenchymal stem cells were used. The cells were derived from the ATCC standard cell bank (PCS-500-010, Lot 63739206, ATCC), and the cells were used in all the following experiments.

[0060] 1. Experimental method

[0061] 1. Preparation of culture medium:

[0062] Prepare following culture medium according to the technical method of embodiment 1

[0063]

[0064]

[0065] 1. Experimental methods and results

[0066] 1. Cell culture

[0067] Human umbilical cord 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 pretreated by coating. Eight groups were set up in the experiment, M1, M2, M3, M4, M5, M6, M7 were used respectively, and the culture medium of the control group was medium containing bovine serum (refer to Example 2).

...

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Abstract

The invention relates to a mesenchymal stem cells serum-free culture medium. The serum-free culture medium comprises a basal culture medium and an additive component, wherein the additive component comprises the following components: rhEGF, rhVEGF, rhPDGF-BB, rhFGF-b, rhTGF-beta1, human serum albumin, recombinant deferritin, a recombinant human insulin growth factor and hydrocortisone. The invention also provides an application of the mesenchymal stem cell serum-free medium. The composition is clear and stable; the instability of serum culture is eliminated, and meanwhile, all factors have thesynergistic effect, the umbilical cord mesenchymal stem cells can be efficiently amplified, the cell cycle is shortened, the proliferation is rapid, the yield of three generations of cultured cells reaches 1.5 times or more of that of serum culture, the cell consistency is good, and the biological characteristics and differentiation potential of the umbilical cord mesenchymal stem cells can be efficiently maintained due to the synergistic effect of all factors.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a serum-free medium for mesenchymal stem cells and its use. 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 the mesenchymal stem cells, umbilical cord mesenchymal stem cells (Human Umbilical Cord-Derived Mesenchymal Stem Cells, UC-MSCs) derived from u...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N5/0668C12N2500/25C12N2500/90C12N2501/11C12N2501/115C12N2501/135C12N2501/15C12N2501/165C12N2501/39C12N2501/998
Inventor 于宝利孙芳陈旭陈刚李其雷
Owner 苏州依科赛生物科技股份有限公司
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