A preparation method of an oral colon-targeted self-assembly polymer loaded with stem cells
A colon targeting and polymer technology, applied in the field of biomedicine, can solve problems such as adverse reactions, poor targeting, and easy recurrence of the disease, and achieve good biocompatibility, promote regeneration and repair effects
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Embodiment 1
[0043] Specific steps are as follows:
[0044] (1) Cultivation of mesenchymal stem cells: take the isolated and purified mesenchymal stem cells for cell culture, use MSCM medium MSCM (ScienCell cat no 7501), and culture them in an incubator (37°C, 5% CO 2 ), subculture when the cells reach 70%-80%, and collect the cells for later use;
[0045](2) Synthesis of OKGM: Add 5g of KGM powder to 500mL of deionized water, stir to dissolve, add dropwise 10mL of 0.5mol / L sodium periodate aqueous solution, and stir at 40°C in the dark for 4h; then add 10mL of ethylene glycol to the reaction mixture and stir 2h to neutralize unreacted periodate; dialyze the solution for 3 days with a dialysis membrane (MWCO: 12,000-14,000) until no iodate is contained in the dialysate; centrifuge the reaction product at 2500r / min for 20min, and take The supernatant was vacuum freeze-dried to obtain OKGM, and the dried sample was stored in a desiccator for use in the next step;
[0046] (3) MSCs-(HTCC / OK...
Embodiment 2
[0051] Step (1) and (2) are with embodiment 1;
[0052] (3) MSCs-(HTCC / OKGM) n Synthesis of self-assembled polymers: Chitosan quaternary ammonium salt (HTCC) and oxidized konjac glucomannan (OKGM) were dissolved in PBS (pH7.2) with a final concentration of 0.2mg / mL. Mix 3 mL cationic polymer chitosan quaternary ammonium salt solution with mesenchymal stem cells in a shaker at 37° C., 100 rpm constant rotation for 30 min, and centrifuge to obtain MSCs-HTCC polymer. Afterwards, 3 mL of the anionic polymer oxidized konjac glucomannan aqueous solution and MSCs-HTCC were mixed in a shaker at 37 °C and 100 rpm for 30 min to obtain the MSCs-HTCC / OKGM self-assembled polymer. Repeat the above steps 1 time to get MSCs-(HTCC / OKGM) 2 Self-assembled polymer, 2 is the number of wrapping layers, and HTCC / OKGM is 1 composite layer.
[0053] After testing, polymers exposed to simulated gastric juice or simulated bile salt solution at 37°C can prevent acid and bile salt erosion for 2 hours, ...
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