Serum-free medium for culturing mesenchymal stem cells and preparation method of serum-free medium
A serum-free medium and stem cell technology, applied in the field of stem cells, can solve the problems of poor safety and unsuitable application of the medium, and achieve the effect of reducing allergens, avoiding the interference of different concentrations and location components, and being highly targeted.
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Embodiment 1
[0020] This example provides a serum-free medium for cultivating mesenchymal stem cells, the formula and preparation process of which are as follows:
[0021] 1. Formula
[0022]
[0023]
[0024] 2. Preparation process
[0025] 1) Preparation of stem cell growth factor
[0026] Take human amniotic mesenchymal stem cells at P2, dilute them with low-sugar DMEM containing 10% FBS, and then press 1×10 5 ~10×10 5 The cell density of cell / mL is seeded in culture flasks for large-scale culture.
[0027] When the growth confluence of the cells reaches more than 80%, the cells are washed twice with PBS, and then replaced with 1640 selective medium to culture the cells, and the medium is changed every 3 days, and the culture medium of the cultured cells is collected.
[0028] The culture solution was filtered with a 0.45 μm filter membrane to remove large particles to obtain a primary filtrate. Filter the primary filtrate with a 0.22 μm filter membrane to remove small partic...
Embodiment 2
[0033] This example provides a serum-free medium for cultivating mesenchymal stem cells, the formula and preparation process of which are as follows:
[0034] 1. Formula
[0035]
[0036] 2. Preparation process
[0037] 1) Preparation of stem cell growth factor
[0038] Take human amniotic mesenchymal stem cells at P2, dilute them with low-sugar DMEM containing 10% FBS, and then press 1×10 5 ~10×10 5 The cell density of cell / mL is seeded in culture flasks for large-scale culture.
[0039] When the growth confluence of the cells reaches more than 80%, the cells are washed twice with PBS, and then replaced with 1640 selective medium to culture the cells, and the medium is changed every 3 days, and the culture medium of the cultured cells is collected.
[0040] The culture solution was filtered with a 0.45 μm filter membrane to remove large particles to obtain a primary filtrate. Filter the primary filtrate with a 0.22 μm filter membrane to remove small particles below 3K...
Embodiment 3
[0045] This example provides a serum-free medium for cultivating mesenchymal stem cells, the formula and preparation process of which are as follows:
[0046] 1. Formula
[0047]
[0048] 2. Preparation process
[0049] 1) Preparation of stem cell growth factor
[0050] Take human amniotic mesenchymal stem cells at P2, dilute them with low-sugar DMEM containing 10% FBS, and then press 1×10 5 ~10×10 5 The cell density of cell / mL is seeded in culture flasks for large-scale culture.
[0051] When the growth confluence of the cells reaches more than 80%, the cells are washed twice with PBS, and then replaced with 1640 selective medium to culture the cells, and the medium is changed every 3 days, and the culture medium of the cultured cells is collected.
[0052] The culture solution was filtered with a 0.45 μm filter membrane to remove large particles to obtain a primary filtrate. Filter the primary filtrate with a 0.22 μm filter membrane to remove small particles below 3K...
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