Double layer artificial skin and preparation method thereof
An artificial skin, double-layer technology, applied in the field of double-layer artificial skin and its preparation, can solve the problems of unfavorable wound epidermal cells crawling wound surface, difficult to observe wound infection, increase immune rejection, etc., to reduce scar hyperplasia, Prevent wound infection, good biocompatibility effect
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Embodiment 1
[0019] Step 1). Acquisition of fibroblasts: Take the foreskin tissue after neonatal resection, wash it with phosphate buffer saline (PBS) after disinfection, remove the subcutaneous fat and muscle layer, cut the skin into strips, and use 4U / ml medium Protease solution for digestion for 2 hours; remove the epidermis, cut the dermal tissue into pieces, digest with 625U / ml collagenase solution for 2.5 hours, and harvest fibroblasts for culture; use cell factory or spinner bottle for the primary cultured fibroblasts Cultivation systems, or methods such as bioreactors to expand cultivation.
[0020] Step 2). Preparation of decellularized small intestinal submucosa: Rinse fresh porcine jejunum with distilled water, cut into required segments, soak and sterilize in an aqueous solution containing 0.2% peracetic acid and 10% ethanol for 1 hour, and rinse with PBS solution Rinse; mechanically scrape off the small intestinal mucosa, muscle layer and serosa, rinse with PBS to obtain the s...
Embodiment 2
[0025] Step 1). Acquisition of fibroblasts: refer to the literature Dermal matrix as a carrier for in vivo delivery of human adipose-derived stem cells. Biomaterials. 2008 Apr; 29(10): 1431-42. It has been proved that adipose-derived mesenchymal stem cells can Differentiate into fibroblasts. Take healthy adipose tissue discarded during the operation, wash it with phosphate buffered saline (PBS) after disinfection, cut up the adipose tissue, digest it with 625U / ml collagenase solution for 150 minutes, and inoculate the harvested adipose-derived mesenchymal stem cells into culture bottles Cultivate; and then use the method of cell factory, or spinner bottle culture system, or bioreactor to expand the culture.
[0026] Step 2). Decellularized dermis preparation: rinse the fresh pig skin with distilled water, remove the epidermis and subcutaneous tissue layer mechanically, cut into 0.4mm thick, 5cm 2 Size, soak and disinfect in an aqueous solution containing 0.2% (v / v) peracetic ...
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