Mesenchymal stem cell culture kit
A stem cell and cell culture technology, 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] Embodiment 1: the preparation of kit
[0024] 1. Preparation of components
[0025] Preparation of cell washing solution: Dissolve 9 g of NaCl in deionized water and adjust the volume to 1000 ml with deionized water, autoclave it and place it in a refrigerator at 4°C for later use.
[0026] Preparation of cell culture medium A: dissolve cytokine LIF, cytokine bFGF and compound Cephaloziellin N 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 CephaloziellinN was 25 μg / ml. Store in a 4°C refrigerator for later use.
[0027] Cell culture medium B is sterile fetal bovine serum, which is stored in a -20°C refrigerator for later use.
[0028] Cell digestion solution: dissolve Collagenase II in deionized water so that the concentration of Collagenase II is 0.2g / 100ml, and store in a 4°C refrigerator for later use.
[0029] 2. Packing of components ...
Embodiment 2
[0031] Example 2: Preparation of the kit, compared with Example 1, only the cell culture solution A does not add CephaloziellinN
[0032] 1. Preparation of components
[0033] Preparation of cell washing solution: Dissolve 9 g of NaCl in deionized water and adjust the volume to 1000 ml with deionized water, autoclave it and place it in a refrigerator at 4°C for later use.
[0034] Preparation of cell culture solution A: cytokine LIF and cytokine bFGF were dissolved in sterile Knockout-DMEM medium (liquid), the concentration of cytokine LIF was 100 ng / ml, and the concentration of cytokine bFGF was 100 ng / ml. Store in a 4°C refrigerator for later use.
[0035] Cell culture medium B is sterile fetal bovine serum, which is stored in a -20°C refrigerator for later use.
[0036] Cell digestion solution: dissolve Collagenase II in deionized water so that the concentration of Collagenase II is 0.2g / 100ml, and store in a 4°C refrigerator for later use.
[0037] 2. Packing of compone...
Embodiment 3
[0039] Example 3: Isolation and rapid expansion of umbilical cord mesenchymal stem cells
[0040] The kits prepared in Example 1 (experimental group) and Example 2 (control group) were used for the isolation and rapid expansion of umbilical cord mesenchymal stem cells.
[0041] Preparation of cell culture medium: Mix cell culture medium A and cell culture medium B at a volume ratio of 5:1.
[0042] 1. Aseptically collect the isolated umbilical cord (the source is human), immerse in the cell culture medium containing double antibody (the cell culture medium containing double antibody is composed of cell culture medium, penicillin and streptomycin, the concentration of penicillin is 1g / 100ml, The concentration of streptomycin is 1g / 100ml), and the glass container is sealed and transported to the laboratory.
[0043] 2. Cut the umbilical cord about 10 cm long in the biological safety cabinet, wash the blood stains with cell washing solution; remove the outer amnion, 2 umbilical ...
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