Application of PGE2-energized mesenchymal stem cell preparation in preparation of medicine for treating lung injury
A technology for stem cells and lung injury, applied in the field of medicine, can solve the problems of limited source of donors and high costs, and achieve the effect of reducing lung injury, significant treatment effect, and reducing lung injury.
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[0025] The preparation method of the PGE2 energized mesenchymal stem cell preparation of the present invention includes: mixing PGE2 and mesenchymal stem cells to obtain the PGE2 energized mesenchymal stem cell preparation.
[0026] In the present invention, the culturing time is preferably 12 to 24 hours, more preferably 12 to 18 hours, and more preferably 12 hours. In the present invention, when the PGE2 and mesenchymal stem cells are mixed and cultured, the concentration of PGE2 in the medium is preferably 2-10 μmol / L, more preferably 2-5 μmol / L, and more preferably 2 μmol / L.
[0027] In the present invention, the medium of the mixed culture is based on DMEM / F12 medium, and preferably also includes the following components: fetal bovine serum 100 mL / L, L-glutamine 10 g / L, optional Amino acid solution 10mL / L and penicillin-streptomycin mixture 10mL / L.
[0028] The invention can improve the anti-inflammatory ability of the mesenchymal stem cells by cultivating PGE2 and the m...
Embodiment 1
[0042] PGE2 empowers MSCs and collects:
[0043] Placental mesenchymal stem cells at passages P3-P5 were plated at 5 × 10 per well. 4The cells were seeded in a 6-well plate. After 24 hours, when the cells grew well, they were replaced with complete medium containing PGE2 (the concentration of PGE2 in the complete medium was 2 μmol / L), and treated with PGE2 for 12 hours, and then the cells were digested for Collect; the complete medium is based on DMEM / F12 medium, and only contains components of the following concentrations: fetal bovine serum 100mL / L, L-glutamine 10g / L, non-essential amino acid solution 10mL / L and penicillin-streptomycin mixture 10mL / L. The morphology of PGE2-energized MSCs was observed under an optical microscope, and the results were as follows figure 1 shown.
[0044] Depend on figure 1 It can be seen that there is no significant difference in the morphology of MSCs before and after energization, indicating that PGE2 energization has no significant effe...
Embodiment 2
[0046] Methods of extracting exosomes (EVs) from PGE2-enhanced MSCs:
[0047] 1) Preparation of exosome-free serum: fetal bovine serum was placed in an ultra-centrifugation tube, centrifuged at 120,000g at 4°C for 2 hours, and the supernatant (exosomes were precipitates) was extracted in a clean bench, and filtered with a 0.22 μm filter. Filter the supernatant and store it in a -80°C refrigerator for use;
[0048] 2) Placental mesenchymal stem cells at passages P3~P5 were prepared at 3 × 10 per flask. 6 The cells were inoculated into T75 culture flasks. After 24 hours, when the cells were in a good growth state, they were replaced with the complete medium containing PGE2 described in Example 1. After PGE2 treatment for 12 hours, the growth was in the logarithmic growth phase, and the cell fusion reached At 80%, the complete medium was removed and washed twice with PBS. After washing, the conditioned medium was added to the culture flask for cultivation. After culturing for 24...
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