Double-cell co-packaging microgel for pancreas islet transplantation and preparation method of double-cell co-packaging microgel
A technology of islet transplantation and islet cells, which is applied in capsule delivery, pharmaceutical formulation, medical science, etc., can solve the problems of immune system rejection, decreased activity of islets, etc., and achieve mild reaction conditions, prolong survival and function maintenance, and good material transfer efficiency Effect
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[0028] The preparation of the gelatin modified by the carbon-carbon double bond used below includes the following process:
[0029] Weigh 5g of gelatin and dissolve it in 50mL of PBS buffer. After stirring and dissolving at 60°C for 1 hour, the temperature was lowered to 50°C in a water bath, and 10 mL of methacrylic anhydride was added dropwise at a rate of 0.5 mL / min with a syringe pump, and reacted for 3 hours. When it was slightly cooled to 40° C., the reactant was diluted with 250 mL of warm PBS at the same temperature to terminate the reaction. After cooling to room temperature, put it into a dialysis bag with a molecular weight cut-off of 12,000-14,000 Da, seal it, and dialyze it in constant-temperature deionized water at 40°C for 7 days. The dialyzed solution is centrifuged at 10000-12000 rpm for 10 minutes, repeated 2-3 times, and the supernatant is freeze-dried to obtain carbon-carbon double bond modified gelatin.
[0030] The preparation of the hyaluronic acid mod...
Embodiment 1
[0033] 1) Put 10 mg of carbon-carbon double bond modified gelatin in advance for UV sterilization overnight in a biological safety cabinet, then add it to 1 mL of LPBS solution, heat up to 40°C and stir to dissolve, then add 0.5 mg of photoinitiator phenyl-2, Lithium 4,6-trimethylbenzoylphosphonate (Lap), resulting in an aqueous dispersed phase.
[0034] 2) Dilute the 2% Fluo-Surf 7500 solution five times with fluorinated oil 7500 to a concentration of 0.05 wt%, as the oily continuous phase.
[0035] 3) The two phases are respectively introduced into the T-shaped microfluidic chip. Adjust and control the flow rate of the dispersed phase to 0.1mL / h and 1mL / h, obtain water-in-oil microemulsion droplets by shearing, collect the prepared droplets and 7500 containing surfactant in a 15mL centrifuge tube, and use UV After irradiating with light for 0.5 minutes, a cross-linked cell-loaded microgel was obtained. Use a pipette gun to transfer the microgel and a small part of fluorina...
Embodiment 2
[0037] 1) Put 100mg of carbon-carbon double bond modified gelatin in a biological safety cabinet for ultraviolet sterilization overnight, then add it to 1mL of LPBS solution, heat up to 40°C and stir to dissolve, then add 5mg of photoinitiator phenyl-2,4 , Lithium 6-trimethylbenzoyl phosphonate (Lap) to obtain an aqueous dispersed phase.
[0038] 2) 2% Fluo-Surf 7500 solution was diluted five times with fluorinated oil 7500 to a concentration of 0.4wt%, as the oily continuous phase.
[0039] 3) The two phases are respectively introduced into the T-shaped microfluidic chip. Adjust and control the flow rate of the dispersed phase to 0.6mL / h and 6mL / h, obtain water-in-oil microemulsion droplets by shearing, collect the prepared droplets and 7500 containing surfactant in a 15mL centrifuge tube, and use UV Light was irradiated for 3 minutes to obtain a cross-linked cell-loaded microgel. Use a pipette gun to transfer the microgel and a small part of fluorinated oil 7500 to a watch...
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