Tissue engineered cartilage graft and preparation method thereof
A technology of tissue engineering and grafts, applied in tissue regeneration, medical science, prosthesis, etc., can solve the problems that hyaline cartilage cannot be completely repaired, and can not be satisfied at the same time, so as to optimize the secretion of cartilage matrix, low cost, and simple and easy method line effect
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[0030] On the other hand, the embodiment of the present invention provides a preparation method of tissue engineered cartilage graft. This preparation method comprises the steps:
[0031] S01: Obtain the umbilical cord mesenchymal stem cell suspension and the three-dimensional scaffold material respectively, and the three-dimensional scaffold material is prepared from type I collagen and type II collagen;
[0032] S02: Inoculate the above-mentioned umbilical cord mesenchymal stem cell suspension into a three-dimensional scaffold material for absorption to obtain a cell-scaffold complex, and initially incubate the cell-scaffold complex at a constant temperature for 3-5 hours;
[0033] S03: Place the cell-scaffold complex after the initial constant temperature incubation in a rotary bioreactor and incubate at a constant temperature for 3-7 days to obtain tissue engineered cartilage grafts.
[0034]The preparation method of the above-mentioned tissue engineered cartilage graft i...
Embodiment 1
[0053] Example 1 Isolation and culture of umbilical cord mesenchymal stem cells
[0054] 1. Provide umbilical cord mesenchymal stem cell samples (separated by Wharton's glue).
[0055] Prepare three sets of autoclaved instruments (each set is equipped with a pair of scissors and a pair of tweezers). The umbilical cord sample was transferred to a 10cm petri dish and rinsed twice with sterile P / S saline. A syringe is inserted into a blood vessel to flush blood from the umbilical cord. Cut the umbilical cord into small pieces of 2-3cm, and wash the blood in the blood vessels respectively. Cut the umbilical cord along the artery, and then remove two arteries and one vein respectively. Take Wharton glue (avoid the skin), cut to 1-2mm 3 size, evenly pasted on the bottom of a 10cm culture dish (with an interval of about 1cm), then add 2-3mL medium until all tissue pieces are wetted, and store at 37°C, 5% CO 2 Cultivate overnight under conditions, continue to add medium to cover ...
Embodiment 2
[0060] Example 2 Preparation of three-dimensional scaffold material
[0061] A method for preparing a three-dimensional scaffold material, comprising the steps of:
[0062] S111: Obtain 0.5 g of type I collagen and 0.5 g of type II collagen raw materials.
[0063] S112: dissolving the above-mentioned type I collagen and type II collagen in water to obtain a mixed solution, and placing the mixed solution in a mold for the first freezing and the first vacuum drying to obtain an initial scaffold material. The specific process is as follows:
[0064] Dissolve the above type I collagen in 10mL of deionized water, then stir overnight to completely dissolve the collagen; add the above type II collagen into the I collagen solution, mix well overnight to obtain a mixture, and put it in 4°C Store in the refrigerator for 24-48 hours to remove air bubbles. Draw up the prepared mixture with a 5mL needle, inject 1mL of the mixture into a well of a 24-well plate, and freeze overnight (24h...
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