A kind of exosome freeze-dried powder and its preparation method and preparation comprising the exosome freeze-dried powder
A technology of exosomes and freeze-dried powder, applied in prostheses, pharmaceutical formulations, bone/connective tissue cells, etc., can solve the problems of poor filling effect, large immune rejection, short maintenance time, etc., to achieve convenient storage, promote Effects of cell differentiation and regeneration and lowering glass transition temperature
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Embodiment 1
[0026] A kind of exosome freeze-dried powder, its preparation method comprises the following steps:
[0027] (1) Preparation of platelet-rich plasma: take autologous peripheral blood, centrifuge at 800g / min for 6min, then absorb the middle part, freeze it at -40°C for 3h, take it out and freeze-thaw it at 2000rpm / min Centrifuge for 6 minutes, remove the solid debris at the bottom, and obtain platelet factor-rich plasma, which is set aside;
[0028] (2) Preparation of exosomes: Absorb autologous fat to prepare P3-generation adipose-derived mesenchymal stem cells, culture P3-generation adipose-derived mesenchymal stem cells, and wash the cells with normal saline when the degree of cell confluence reaches 75%. Then the cells were transferred to the serum-free medium and cultured for 20 hours, the cell culture supernatant was collected, centrifuged at 2°C and 800g centrifugal force for 10min, discarded the precipitate, and then carried out ultrasonography under the condition of th...
Embodiment 2
[0033] A kind of exosome freeze-dried powder, its preparation method comprises the following steps:
[0034] (1) Preparation of platelet-rich plasma: take autologous peripheral blood, centrifuge at 2000g / min for 12min, then absorb the middle part, freeze it at -80°C for 8h, take it out and freeze-thaw it at 4000rpm / min Centrifuge for 10 minutes, remove the solid debris at the bottom, and obtain platelet factor-rich plasma, which is set aside;
[0035] (2) Preparation of exosomes: Absorb autologous fat to prepare P6-generation adipose-derived mesenchymal stem cells, culture P6-generation adipose-derived mesenchymal stem cells, and wash the cells with normal saline when the degree of cell confluence reaches 85%. Then the cells were transferred to serum-free medium and cultured for 30 hours, and the cell culture supernatant was collected, centrifuged at 6°C and 1200g centrifugal force for 20 minutes, discarded the precipitate, and then subjected to ultrasonography under the condi...
Embodiment 3
[0040] A kind of exosome freeze-dried powder, its preparation method comprises the following steps:
[0041] (1) Preparation of platelet-rich plasma: take autologous peripheral blood, centrifuge at 1500g / min for 10min, then absorb the middle part, freeze it at -80°C for 6h, take it out and freeze-thaw it at 3000rpm / min Centrifuge for 8 minutes, remove the solid debris at the bottom, and obtain platelet factor-rich plasma, which is set aside;
[0042] (2) Preparation of exosomes: Absorb autologous fat to prepare P4 adipose-derived mesenchymal stem cells, culture P4 adipose-derived mesenchymal stem cells, and wash the cells with normal saline when the degree of cell confluence reaches 80%. Then the cells were transferred to serum-free medium and cultured for 24 hours, and the cell culture supernatant was collected, centrifuged at 4°C and 1000g centrifugal force for 15 minutes, discarded the precipitate, and then subjected to ultrasonography under the condition of a current carry...
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