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
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[0027] Example 1
[0028] The serum-free medium for mesenchymal stem cells provided in this example consists of the following components: a basal medium and additional components, the basal medium is DMEM / F12, and the additional components are denoted as MAX1 component, MAX1 component 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 (all components can be purchased from Sigma, Aladdin and other reagent companies) and the content of each component (based on the volume of the DMEM / F12 component):
[0033]
[0034]
[0035] In this example, the basic medium DMEM / F12 can also be purchased directly from the reagent company.
[0036] In this example, the basal medium DMEM / F12 is made of DMEM / F12 powder (Sigma, D2906), water and 1.2 g of sodium bicarbonate (S...
Example Embodiment
[0038] Example 2
[0039] This example provides verification of the effect of the serum-free medium for mesenchymal stem cells of Example 1.
[0040] In the experiment 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 these 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 a 100mm cell culture dish. 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) plus 10% fetal bovine serum (ExCellBio, FND500)) for continuous culture, the amount added is about 15ml , Every 48 ~ Change the medium on...
Example Embodiment
[0058] Example 3
[0059] This example provides verification of the effects of various culture media. In the experiment 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 these cells were used in all the following experiments.
[0060] 1. Experimental method
[0061] 1. Preparation of culture medium:
[0062] According to the technical method of Example 1, the following medium was prepared
[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 a 100mm cell culture dish, and all the cell culture dishes used in the experiment were coated and pretreated. The experiment set up 8 groups, respectively using M1, M2, M3, M4, M5, M6, M7, and the control group used a medium containing bovine serum (refer to Example 2).
[0068] Add the above medium for continuo...
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