Stem cell exosome preparation as well as preparation method and application thereof
An exosome and stem cell technology, applied in the biological field, can solve the problems of general treatment effect, inability to fundamentally cure the disease, and large patient trauma, and achieve the effects of good curative effect, shortened time, and convenient preparation of preparations.
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Embodiment 1
[0045] Example 1 Preparation of Dental Pulp Stem Cell Exosomes
[0046] Primary culture of dental pulp stem cells: Take the wisdom teeth of healthy people and perform aseptic cleaning in an ultra-clean bench. After cleaning, cut the teeth with forceps and take out the pulp; cut the pulp with surgical scissors, and use type I collagenase Digest with neutral protease for 30min, centrifuge at 1500rpm for 10min, discard the supernatant, resuspend with DMEM+10%FBS complete medium, inoculate into 6-well plate, transfer to 5%CO 2, Cultivate in an environment of 37°C; discard the supernatant after 5 days of culture, supplement with new complete medium and continue to culture, and the cells can be passaged when they form larger clones;
[0047] Subculture of dental pulp stem cells: Take subcultured P0 cells, discard the original culture medium in the dish, add 10mL PBS buffer solution to gently wash the cell growth surface; discard the PBS washing solution, add 2mL 0.25% trypsin, and s...
Embodiment 2
[0053] Example 2 Preparation of exosome preparation
[0054] Add human serum albumin to normal saline to prepare a human serum albumin solution with a mass concentration of human serum albumin of 5%; then add the dental pulp stem cell exosomes and dental pulp prepared in Example 1 to the human serum albumin solution Stem cells, so that each 5mL solution contains 5×10 6 dental pulp stem cells and from 5×10 6 The exosomes extracted from dental pulp stem cells were prepared and stored at 4°C.
Embodiment 3
[0055] Example 3 Preparation of exosome preparation
[0056] Add human serum albumin to normal saline to prepare a human serum albumin solution with a mass concentration of human serum albumin of 2%; then add the dental pulp stem cell exosomes prepared in Example 1 and Dental pulp stem cells, so that each 5mL solution contains 5×10 6 dental pulp stem cells and from 1×10 6 The exosomes extracted from dental pulp stem cells were prepared and stored at 4°C.
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