Arthrodial cartilage proto micro-carrier
A technology of articular cartilage and microcarriers, which is applied in the field of biomedical tissue engineering, can solve the problems of incomplete separation of cells and microcarriers, affecting the quality of engineered cartilage, and damage to chondrocytes, achieving cheap reagents, avoiding loss, and growing state Good results
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Embodiment 1
[0024] 1. Take 1kg of fresh pig articular cartilage, freeze-dry it, pulverize it with a pulverizer, and sieve to obtain particles with a diameter of 200μm for use;
[0025] 2. Add 10 times the volume of 0.25% trypsin to the granules, keep shaking at 37°C for 48h and then wash with distilled water 3 times, use 1% Triton X-100 at 4°C to shake for 72h, then rinse with distilled water for 48h;
[0026] 3. Centrifuge at 1000 rpm for 10 minutes, take the precipitate, mix the precipitate with distilled water in a volume of 1:3, and put it in a freeze dryer for 12 hours to freeze-dry it. After 25kGy cobalt 60 Sterilized by irradiation, digested with 10 times the volume of 0.25% trypsin for 15 minutes, stirred, then pulverized into granules, and washed with distilled water 3 times to obtain.
Embodiment 2
[0028] 1. Take 4kg of fresh bovine joint cartilage, freeze-dried, crush it with a pulverizer, and sieve to obtain particles with a diameter of 150μm for use;
[0029] 2. Add 10 times the volume of 0.25% trypsin (purchased from Sigma) to the granules, continue shaking at 37°C for 48h, and then wash 3 times with distilled water, then use 1% Triton X-100 at 4°C to continue shaking for 72h, then Rinse with distilled water for 48h;
[0030] 3. Centrifuge at 1000 rpm for 10 minutes, take the precipitate, mix the precipitate with distilled water in a volume of 1:3, and put it in a freeze dryer for 12 hours to freeze-dry it. After 25kGy cobalt 60 Sterilized by irradiation, digested with 10 times the volume of 0.25% trypsin (purchased from Sigma) for 15 minutes, stirred, pulverized into granules, and washed with distilled water 3 times to obtain.
Embodiment 3
[0032] 1. Take 0.5kg of fresh human articular cartilage, freeze-dried, crush it with a pulverizer, and sieve to obtain particles with a diameter of 180μm for use;
[0033] 2. Add 10 times the volume of 0.25% trypsin (purchased from Sigma) to the granules, continue shaking at 37°C for 48h, and then wash 3 times with distilled water, then use 1% Triton X-100 at 4°C to continue shaking for 72h, then Rinse with distilled water for 48h;
[0034] 3. Centrifuge at 1000 rpm for 10 minutes, take the precipitate, mix the precipitate with distilled water in a volume of 1:3, and put it in a freeze dryer for 12 hours to freeze-dry it. After 25kGy cobalt 60 Sterilized by irradiation, digested with 0.25% trypsin for 15 minutes, stirred, then pulverized into granules, and washed with distilled water 3 times to obtain.
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