Engineered extracellular matrix preparation method
A technology of tissue engineering and extracellular matrix, applied in the field of artificial organs of biomaterials, can solve the problems of poor clinical effect, achieve the effects of repairing malnutrition, good therapeutic effect, and preventing recurrence
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Embodiment 1
[0046] Now in conjunction with experiment, preparation method of the present invention is described in further detail:
[0047] 1. Preparation of culture medium
[0048] (1) Basal culture medium
[0049] Ingredient Final Concentration
[0050] Commercial minimum essential medium DMEM 67.5% (v / v)
[0051] Commercial culture medium F12 22.5% (v / v)
[0052] Fetal bovine serum 10% (v / v)
[0053] Insulin 10ng / ml
[0054] Hydrocortisone 30ng / ml
[0055] Adenine 0.22mM
[0056] Transferrin 6mg / ml
[0057] Epidermal growth factor 80ng / ml
[0058] Antibiotics 180i.u / ml
[0059] (2) Culture medium 1
[0060] Basal Medium + Bovine Pituitary Extract 0.5% (V / V)
[0061] (3) Culture medium 2
[0062] Basal medium + vitamin C20μg / ml
[0063] (4) Culture medium 3
[0064] Basal medium + basic fibroblast growth factor 5ng / ml
[0065] 2. Extraction of matrix protein: the present invention adopts the matrix protein mixture extracted from the skin of mammals (can be cattle, rabbits...
Embodiment 2
[0071] Embodiment 2. burn wound repair test
[0072] 1) Preparation of animal skin deep second-degree scald model: 50kg Yorkie pigs were anesthetized, and the back skin was prepared for disinfection and sterile towels were spread. Using a scald instrument: temperature 100°C, pressure 1kg, time 25s; prepare the burn wound ( Figure 4 ).
[0073] 2) 24 hours after the preparation of the above model, anesthetize the animal, completely remove all wound necrotic tissue, repair the wound, take out the tissue-engineered extracellular matrix prepared by the present invention from the sterile sealed bag, tear off the nylon film, and cover the wound. Pack it up. The blank control group was covered with conventional Vaseline gauze. The wound was opened for the first time 5 days after the operation, and the dressing was changed routinely. The wound was observed and the wound tissue was observed by HE staining and specific staining, and the average healing time of the wound was calculat...
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