Rapid preparation of accellular porcine intestinal submucosa
A submucosa and decellularization technology, applied in medical science, tissue regeneration, prostheses, etc., can solve the problems of reducing the mechanical properties of the final ECM, incompleteness, cumbersome complexity, etc., to maintain chemical composition and mechanical properties, and ensure compatibility Sexual, non-toxic promoting effect
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[0032] A rapid preparation method of acellular porcine small intestinal submucosa includes the following steps:
[0033] S1: Material pretreatment: Cut the fresh pig jejunum, clean it repeatedly, and then cut it to an appropriate length as needed, generally 10-20cm long; scrape off the serosal layer, mucosal layer and muscle layer of the jejunum with a blunt spatula. Rinse it with deionized water, then place it in the newly prepared pretreatment solution, soak it for 30-60 minutes, and then set it aside.
[0034] S2: One-time decellularization treatment: Rinse the pretreated submucosa of the pig small intestine with deionized water, drain it, transfer it to the cell lysate, sonicate for 10-30 minutes or soak for 30-120 minutes with shaking, and then use it for another 30 minutes. Rinse with deionized water at -45℃ for 10-60 minutes;
[0035] S3: Degreasing treatment: the submucosa of the pig small intestine after a decellularization treatment is rinsed with deionized water, drained,...
Embodiment 1
[0065] 1. Preparation of pretreatment liquid
[0066] Weigh sodium silicate (Na2SiO 3 ) 100g, 60g hydrogen peroxide, add 700ml deionized water and mix well; measure 20-60nm colloidal silver 40mg, then slowly add colloidal silver while stirring the hydrogen peroxide solution, adjust the pH to 2.0 with 1N HCl after addition, Finally, add deionized water to 1 liter to obtain the pretreatment solution for rapid preparation of decellularized porcine intestinal submucosa (p-SIS) of this example.
[0067] 2. Preparation of cell lysate
[0068] Weigh 1g of borax and 10.3g of sodium carbonate, add 700ml of deionized water and mix well; add 200ml of pre-prepared 5M NaCl mother liquor and 40ml of 0.5M EDTA mother liquor, then slowly add 10ml of liquid NP-40(1g / ml), after the addition, adjust the pH to 11 with 1N NaOH, and finally add deionized water to 1 liter to obtain the cell lysate for rapid preparation of decellularized pig small intestinal submucosa (p-SIS) of this example. Each mother...
Embodiment 2
[0086] This example is basically the same as Example 1, except that the rapid preparation method of decellularized porcine small intestinal submucosa (p-SIS) partially omitted step 5) (that is, the step of refolding treatment).
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