Platelet lysate supported micro-sphere preparation method
A technology of platelet lysate and platelet, which is applied in the direction of blood/immune system cells, cells fixed on/in organic carriers, bone/connective tissue cells, etc. It can solve the problem of large consumption of platelet lysate, poor adsorption effect of microcarriers, etc. problem, to achieve the effect of effective proliferation, promotion of attachment, and increase of specific surface area
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specific Embodiment approach 1
[0024] Specific embodiment one: In this embodiment, a method for preparing microspheres loaded with platelet lysate is carried out according to the following steps:
[0025] 1. Add basement membrane proteoglycan solution to the platelet lysate rich in CD41+ and CD81+ microcapsules, and stir evenly while adding to obtain composition A; wherein the platelet lysate and basement membrane protein rich in CD41+ and CD81+ microcapsules The volume ratio of polysaccharide solution is (3.7-5.5):(0.8-2.4);
[0026] 2. At room temperature, add the sodium alginate solution to composition A, and mix well to form composition B; wherein the volume ratio of the sodium alginate solution to the platelet lysate rich in CD41+ and CD81+ microcapsules is (3.0- 5.0):(3.7-5.5);
[0027] 3. Add composition B to a spray gun equipped with a porous nozzle, use nitrogen as the power, and spray into 3-10% CaCl at 4°C with 90rpm magnetic stirring at a speed of 1-30m / s 2 In aqueous solution, microspheres wi...
specific Embodiment approach 2
[0032] Specific embodiment two: the difference between this embodiment and specific embodiment one is: the preparation method of the platelet lysate rich in CD41+, CD81+ microcapsules in step one is:
[0033] 1. Use a focused extracorporeal shock wave instrument with a frequency of 15Hz and an energy-flux density of 0.15-0.20mJ / mm 2 , after shocking platelet-rich plasma or platelet-rich saline 200-500 times, at 37°C CO 2 Incubate in the incubator for 10-15 minutes; then at a frequency of 15Hz, the energy flux density is 0.20-0.25mJ / mm 2 , shock 700-1000 times, at 37°C CO 2 Incubate in the incubator for 20-30 minutes; then at a frequency of 15Hz, the energy flux density is 0.25-0.30mJ / mm 2 , shock 1500-2000 times, at 37°C CO 2 Incubate in an incubator for 10 minutes to obtain a suspension;
[0034] 2. Put the suspension in step 1 into a sealed freezing tube, put it into liquid nitrogen to freeze for 20-40 minutes, take it out and put it in a water bath to shake and melt, re...
specific Embodiment approach 3
[0036] Specific embodiment 3: The difference between this embodiment and specific embodiment 1 or 2 is that the platelet concentration in platelet-rich plasma or platelet-rich saline is (6-15)×10 5 pieces / μl. It is the same as the specific embodiment 1 or 2.
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