Sustained-release microsphere containing immature dendritic cell exosome and preparation method of sustained-release microsphere
A technology of dendritic cells and slow-release microspheres, applied in the biological field, to achieve long-lasting immunosuppression, maintenance of immune tolerance, and good biocompatibility
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[0038] The preparation method of the above-mentioned slow-release microspheres containing immature dendritic cell exosomes comprises the following steps:
[0039] Step 1, preparation of immature dendritic cell exosomes:
[0040] S1 Obtain bone marrow dendritic cells and culture them in the medium containing granulocyte-macrophage colony-stimulating factor and interleukin-4;
[0041] After the S2 culture is completed, the cell culture fluid is collected and then subjected to differential ultracentrifugation;
[0042] After S3 centrifugation, discard the supernatant, resuspend the pellet with PBS, then centrifuge again, discard the supernatant, and resuspend the pellet with PBS to obtain immature dendritic cell exosomes;
[0043] Step 2, preparation of slow-release microspheres:
[0044] S1 Take the immature dendritic cell exosome solution and the sodium alginate solution according to the volume parts, and stir well to obtain the exosome sodium alginate suspension;
[0045] S...
Embodiment 1
[0057] This example provides a slow-release microsphere containing immature dendritic cell exosomes, the preparation method of which includes the following steps:
[0058]1. Cell culture and preparation of exosomes: Bone marrow dendritic cells (BMDCs) were obtained in a culture medium containing granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) Cultivate, change the medium in half the amount every other day, and observe the cell state under the optical electron microscope on the 7th day of culture (such as figure 1 ), and the cell culture fluid was collected, and imDCex was obtained by differential ultracentrifugation. Differential ultracentrifugation method: centrifuge the culture medium at 300g for 10min to remove cells; centrifuge at 2000g for 10min to remove dead cells; centrifuge at 10000g for 30min to remove cell debris; centrifuge at 100000g for 70min and discard the supernatant , the pellet was resuspended with phosphate buffered salin...
Embodiment 2
[0064] This example provides a slow-release microsphere containing exosomes from immature dendritic cells. The culture of the cells and the preparation of the exosomes are the same as in Example 1. The preparation process of the slow-release microspheres is as follows:
[0065] Take 6 parts of immature dendritic cell exosome solution and 97 parts of 0.8% sodium alginate solution according to the volume fraction, stir well and evenly to obtain exosome sodium alginate suspension; put the exosome sodium alginate suspension into Into the electrostatic sprayer, the operating parameters of the electrostatic sprayer are set to be: the voltage is 14kv, the speed of the injection pump is 0.7ml / min, and it is started, and the spray droplets are sprayed from the nozzle and dispersed in the solidified liquid, and the buffer is obtained. Release the microspheres, and then observe under the light microscope, the results are as follows Figure 6 .
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