Mesenchymal stem cell nutrient solution
A mesenchymal stem cell and culture medium technology, applied in the field of mesenchymal stem cell culture, can solve the problems of high cost, cell pollution, and complex formula of serum-free culture medium, and achieve the effect of high safety and simple formula
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Embodiment 1
[0021] Wash 20cc adipose tissue 3 times with phosphate buffered saline, add collagenase (Type II, 50-100U / ml) to digest the tissue for one hour, centrifuge the digest and wash the resulting cells 3 times with phosphate buffered saline; mix the cells 7ml of culture solution was placed in a T-75 cell culture flask for cultivation. The components of the culture solution in this example were: alpha-MEM medium, B27 serum-free additive with a volume ratio of 1%, bFGF at 10 ng / ml, bFGF at 10 ng / ml EGF, 0.5mg / ml fetuin, 100units / ml each of penicillin and streptomycin, use the above culture medium, replace the culture medium once every 3 days, and detect when the cells grow to a T-75 cell culture flask. No less than one million adipose-derived mesenchymal stem cells were cultured.
Embodiment 2
[0023] Use an ethanol solution with a volume ratio of 70% to disinfect the oral cavity of the human body, and then use a 2mm diameter biopsy instrument to extract a tissue sample (not less than 2mmx2mmx2mm) from the human gum, wash it with phosphate buffer for 3 times, and add collagenase (Type II, 50-100U / ml) to digest the tissue for one hour, centrifuge the digest and wash the resulting cells 3 times with phosphate buffered saline; mix the gingival tissue cells with 10ml culture solution and put them in a T-75 cell culture bottle for cultivation. The components of the culture medium are: DMEM / F12 serum-free medium, B27 serum-free supplement with a volume ratio of 3%, bFGF at 50ng / ml, EGF at 50ng / ml, fetuin at 2mg / ml, and penicillin at 150units / ml each and streptomycin, use the above culture medium, replace the culture medium once every 3 days, when the cells grow to a T-75 cell culture flask, it is detected that no less than one million gingival tissue mesenchymal stem cells ...
Embodiment 3
[0026] Thaw adipose-derived mesenchymal stem cells (not less than 400,000 cells) to 37 degrees, wash the obtained cells twice with phosphate buffer, mix the cells with 20ml of culture medium and put them in a T-150 cell culture bottle for cultivation. This implementation The culture solution components of the example are: alpha-MEM medium, B27 serum-free supplement with a volume ratio of 2%, bFGF at 20ng / ml, EGF at 20ng / ml, fetuin at 1mg / ml, penicillin at 100units / ml each And streptomycin, using the above culture medium, replace the culture medium once every 3 days, when the cells grow to a T-75 cell culture flask, it is detected that no less than 3 million adipose-derived mesenchymal stem cells have been cultured.
[0027] Compared with the culture of adipose-derived mesenchymal stem cells using the serum-free medium formula of Dromard C et al., 2011, the cells will grow in suspension in the culture flask, while the mesenchymal stem cells of this example can be attached to the...
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