A kind of hollow open gelatin cell microcarrier and its preparation method and application
A technology of cell microcarriers and microcarriers, applied in prosthetics, medical science, etc., can solve the problems of insufficient space reserved for newborn tissues, unfavorable high-density delivery of therapeutic cells, and low efficiency of delivery of cells, and achieve good biodegradability and cell compatibility, excellent biocompatibility, and good application prospects
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Embodiment 1
[0060] Embodiment 1 Preparation of hollow open gelatin cell microcarriers of the present invention
[0061] 1. Preparation method
[0062] S1. Preparation of gelatin and beeswax microspheres by double emulsion method
[0063] S11. Weigh 4mg of gelatin, dissolve in 40mL of pure water, and heat to 80°C;
[0064] S12. Add 4 mg of beeswax to the gelatin solution of S11 at a rotational speed of 700 rpm, and stir for 5 minutes at a rotational speed of 700 rpm;
[0065] S13. Pour the solution obtained in S12 into edible peanut oil preheated at 60°C, and turn it at 400rpm for 15min;
[0066] S14. Pour the solution obtained in S13 into -20°C pre-cooled ice ethanol, let it stand for 15 minutes, then wash it with dioxane / acetone solution three times, put it into a 60°C oven for drying, and obtain gelatin beeswax microspheres;
[0067] The morphology of the obtained gelatin beeswax microspheres observed under the light microscope is as attached figure 1 shown.
[0068] S2. Gelatin be...
Embodiment 2
[0083] Example 2 Layer-by-layer Self-Assembly Modified Surface Biocompatibility of Hollow Open Gelatin Microcarriers
[0084] In this example, the surface biocompatibility of the hollow open gelatin cell microcarriers prepared in Example 1 was modified to further improve the surface biocompatibility of the microcarriers.
[0085] 1. Weigh 80 mg of hollow open gelatin cell microcarriers, place them in 75% ethanol solution, immerse and sterilize for 4 hours, wash with sterile PBS three times, and place them in a 12-well plate;
[0086] 2. Prepare a mixed solution containing 0.2 mg / mL casein and 4 mg / mL heparin as a negatively charged macromolecule assembly solution, and prepare a mixed solution containing 1.0 mg / mL collagen and 10 g / mL basic fibroblast growth factor as a positively charged macromolecule Assembly solution;
[0087] 3. Add 4mL casein / heparin solution to the 12-well plate where the microcarriers are placed, soak for 15 minutes, so that a layer of casein / heparin ma...
Embodiment 3
[0090] Example 3 Inoculation of bone marrow mesenchymal stem cells with hollow open gelatin microcarriers
[0091] 1. Method
[0092] (1) Place the hollow open gelatin microcarrier obtained in Example 2 in a centrifuge tube (EP tube);
[0093] (2) Digest P3 generation bone marrow mesenchymal stem cells with 0.25% trypsin, count the cells, add 0.5mL of 500,000 cells / mL cell suspension into the EP tube with hollow open gelatin microcarriers, and then Place the EP tube at 37°C, 5% CO 2 Cultivate in the incubator for 8 hours;
[0094] (3) Add 0.5mL of 500,000 cells / mL cell suspension into the EP tube equipped with hollow open gelatin microcarriers, and then place the EP tube at 37°C, 5% CO 2 Cultivate in the incubator for 8 hours;
[0095] (4) Transfer the hollow open gelatin cell microcarriers to a 6-well plate, and use basal medium (α-MEM, 10% FBS, 100 U / mL penicillin, 100 g / mL streptomycin, 0.25 g / mL amphoteric Mycin B) was cultured, and the medium was changed every other ...
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