Artificial skin containing human bone marrow mesenchymal stem cell and its construction method
A bone marrow mesenchymal and artificial skin technology, applied in the field of artificial skin, can solve the problems of toxic hazards in the body, no artificial skin, strong immune antigenicity, etc., achieve good flexibility and enzyme resistance, and induce autologous cell growth , accelerated healing effect
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[0034] Example 1
[0035]The fetal cowhide is depilated, fasciae, fat and blood stained, washed, frozen, cut into 0.5mm thin slices, dehydrated three times with acetone, degreasing three times with petroleum ether, and then vacuumized to remove petroleum ether to make dehydration and degreasing. Fetal cow hides were used as raw materials, 10 g of raw materials were taken, washed with double distilled water, and then added to the reactor, and an aqueous solution prepared by 0.1 g of gastric enzyme and 100 g of water was added. Enzyme treatment was carried out once, under the conditions of 20 ° C, pH=2, mechanical vibration for 24 hours, degraded the covalent bonds between tissue fibers, centrifuged, discarded the supernatant, took the precipitate, washed three times with double distilled water, and added 500 g 0.5M acetic acid aqueous solution, mechanically vibrated for 32 hours at 0 °C, and then added sodium chloride to make the sodium chloride concentration in the solution 2M...
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[0038] Example 2
[0039] The bovine achilles tendon was dehydrated, degreased, and vacuumed to remove petroleum ether according to the method of Example 1, and the dehydrated and degreased bovine achilles tendon was made as a raw material. g trypsin and 100 g of the prepared aqueous solution were subjected to an enzymatic treatment. Under the conditions of 30 °C and pH=6, mechanical vibration was performed for 16 hours to degrade the covalent bonds between tissue fibers. After centrifugation, the supernatant was discarded, and the precipitate was collected. Wash with distilled water four times, add 1000g of 0.5M acetic acid aqueous solution, mechanically vibrate for 28 hours at 10°C, and then add sodium chloride to make the concentration of sodium chloride in the solution reach 2M. After the precipitation is complete, centrifuge and discard the supernatant. , take the precipitate, put it into the dialysis agent and carry out dialysis four times, collect the collagen solution ...
Example Embodiment
[0042] Example 3
[0043] Use the dehydrated and defatted bovine Achilles tendon of Example 2 as a raw material, take 10 g, wash it with double distilled water and add it to the reaction vessel, add the aqueous solution prepared by 0.5 g of pineapple enzyme and 100 g of water to carry out an enzyme treatment, at 40 ° C, pH Mechanical vibration for 8 hours under the condition of =8 to degrade the covalent bonds between tissue fibers, centrifugation, discarding the supernatant, taking the precipitate, washing four times with double distilled water, adding 1500g of 0.5M acetic acid aqueous solution, at 20 ℃ temperature Under mechanical vibration for 24 hours, then add sodium chloride to make the sodium chloride concentration in the solution reach 2M, after the precipitation is complete, centrifuge, discard the supernatant, take the precipitate and put it into a dialysis bag for dialysis three times, collect the collagen solution after dialysis and add In the reactor, an aqueous s...
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