Kit for culturing mesenchymal stem cells
A technology of qualitative stem cells and kits, applied in the field of stem cell culture, can solve the problems of low purity and long primary culture time, and achieve the effect of increasing the passage time
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Embodiment 1
[0023] Example 1: Preparation of the kit
[0024] 1. Preparation of components
[0025] Preparation of cell washing solution: Dissolve 9g of NaCl in deionized water and dilute to 1000ml with deionized water. After autoclaving, place it in a refrigerator at 4°C for later use.
[0026] Preparation of cell culture fluid A: Dissolve cytokine LIF, cytokine bFGF and compound SalpichrolideO in sterile Knockout-DMEM medium (liquid), the concentration of cytokine LIF is 100ng / ml, and the concentration of cytokine bFGF is 100ng / ml , The concentration of the compound SalpichrolideO is 25μg / ml. Place in a refrigerator at 4°C for later use.
[0027] Cell culture medium B is sterile fetal bovine serum, placed in a refrigerator at -20°C for later use.
[0028] Cell digestion solution: Dissolve CollagenaseII in deionized water to make the concentration of CollagenaseII 0.2g / 100ml, and place it in a refrigerator at 4°C for later use.
[0029] 2. Packing of components and preparation of kits
[0030] The...
Embodiment 2
[0031] Example 2: Preparation of the kit. Compared with Example 1, only SalpichrolideO is not added to the cell culture medium A
[0032] 1. Preparation of components
[0033] Preparation of cell washing solution: Dissolve 9g of NaCl in deionized water and dilute to 1000ml with deionized water. After autoclaving, place it in a refrigerator at 4°C for later use.
[0034] Preparation of cell culture fluid A: Dissolve cytokine LIF and cytokine bFGF in sterile Knockout-DMEM medium (liquid), the concentration of cytokine LIF is 100ng / ml, and the concentration of cytokine bFGF is 100ng / ml. Place in a refrigerator at 4°C for later use.
[0035] Cell culture medium B is sterile fetal bovine serum, placed in a refrigerator at -20°C for later use.
[0036] Cell digestion solution: Dissolve CollagenaseII in deionized water to make the concentration of CollagenaseII 0.2g / 100ml, and place it in a refrigerator at 4°C for later use.
[0037] 2. Packing of components and preparation of kits
[0038] The...
Embodiment 3
[0039] Example 3: Isolation and rapid expansion and culture of umbilical cord mesenchymal stem cells
[0040] The kits prepared in Example 1 (experimental group) and Example 2 (control group) were used to separate and rapidly expand umbilical cord mesenchymal stem cells.
[0041] Preparation of cell culture fluid: mix cell culture fluid A and cell culture fluid B in a volume ratio of 5:1.
[0042] 1. Collect the isolated umbilical cord (source from human) aseptically and immerse it in a cell culture medium containing double antibodies (the cell culture medium containing double antibodies consists of cell culture medium, penicillin and streptomycin, and the concentration of penicillin is 1g / 100ml, The concentration of streptomycin is 1g / 100ml), the glass container is sealed and transported to the laboratory.
[0043] 2. Cut the umbilical cord about 10 cm in length in the biological safety cabinet, and wash the blood stains with cell washing solution; remove the outer amniotic membrane,...
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