Collagen melanocyte compound as well as preparation method and application thereof
A technology of melanocytes and collagen, which is applied in the field of biomedicine, can solve the problems of not being suitable for large-scale transplantation, difficulty in fixing the tip, and easy loss of suspension, so as to avoid the loss of a large number of cells, easy to obtain, and simple to prepare scaffolds Effect
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Embodiment 1
[0020] Example 1: Preparation method of cross-linked collagen melanocyte complex
[0021] Cut 1g of collagen sponge into a 1cm*1cm square, put it in a 1000mL beaker, add 500mL of 90% ethanol, stir magnetically at room temperature, add 9.6g of EDC, 11.5g of NHS, and react for 48h. After the reaction, the cross-linked collagen sponge was soaked in 0.1mol / L phosphate buffer solution, stirred by magnetic force for 1 hour, rinsed with purified water, sterilized by irradiation after freeze-drying.
[0022] Cut the sterile collagen sponge into a material of uniform size: the upper surface is 1cm*1cm square, with a thickness of 5mm, and then pre-spread the cut scaffold material on the cell culture plate. Dilute 200μl of melanocytes to 1*10 5 Cells / ml were seeded on collagen scaffolds and placed in a cell culture incubator at 37°C, 5% CO 2 , cultured under saturated humidity conditions. After culturing for 2 hours, add 1ml of cell culture medium (without penicillin and streptomycin)...
Embodiment 2
[0023] Embodiment 2: the preparation method of cross-linked collagen melanocyte complex
[0024] Cut 1g of collagen sponge into a 1cm*1cm square, put it in a 1000mL beaker, add 500mL of 90% ethanol, stir magnetically at room temperature, add 9.6g of EDC, 11.5g of NHS, and react for 48h. After the reaction, the cross-linked collagen sponge was soaked in 0.1mol / L phosphate buffer solution, stirred by magnetic force for 1 hour, rinsed with purified water, sterilized by irradiation after freeze-drying.
[0025] Cut the sterile collagen sponge into a material of uniform size: the upper surface is 1cm*1cm square, with a thickness of 5mm, and then pre-spread the cut scaffold material on the cell culture plate. 400μl melanocytes were divided into 1*10 5 Cells / ml were seeded on collagen scaffolds and placed in a cell culture incubator at 37°C, 5% CO 2 , cultured under saturated humidity conditions. After culturing for 2 hours, add 1ml of cell culture medium (without penicillin and s...
Embodiment 3
[0026] Embodiment 3: the preparation method of cross-linked collagen melanocyte complex
[0027] Cut 1.5g of collagen sponge into a 1cm*1cm square, put it in a 1000mL beaker, add 500mL of 90% ethanol, stir magnetically at room temperature, add 9.6g of EDC, 11.5g of NHS, and react for 48 hours. After the reaction, the cross-linked collagen sponge was soaked in 0.1mol / L phosphate buffer solution, stirred by magnetic force for 1 hour, rinsed with purified water, sterilized by irradiation after freeze-drying.
[0028] Cut the sterile collagen sponge into a material of uniform size: the upper surface is 1cm*1cm square, with a thickness of 5mm, and then pre-spread the cut scaffold material on the cell culture plate. 400μl melanocytes were divided into 1*10 5 Cells / ml were seeded on collagen scaffolds and placed in a cell culture incubator at 37°C, 5% CO 2 , cultured under saturated humidity conditions. After culturing for 2 hours, add 1ml of cell culture medium (without penicilli...
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