Kit and method used for obtaining full liver stent
A kit and liver technology, applied in medical science, tissue regeneration, prosthesis, etc., can solve the problems of incomplete effect and complex method and process, and achieve the effect of strong buffering ability and stable pH environment.
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Embodiment 1
[0026] Example 1. Rat liver decellularization
[0027] 1. Prepare perfusate I and perfusate II:
[0028] Perfusate I: Take 10ml of Triton X-100, dissolve it in 800ml of Tris-HCl with pH 7.5 and 50mM, and dilute to 1000ml to obtain 1% Triton X-100;
[0029] Perfusate II: 10 g of SDS was dissolved in 800 ml of Tris-HCl of pH 7.5 and 50 mM, and the volume was adjusted to 1000 ml to obtain 1% SDS.
[0030] 2. Perfusion to achieve decellularization:
[0031] Insert a cannula from the portal vein of the isolated liver, and perfuse the isolated liver with 1% TritonX-100 perfusion solution at a rate of 10ml / min for 1 hour. At this time, the central area of the liver becomes milky white, and the outflowing perfusate is turbid brown yellow; then Perfuse with double distilled water for 1 hour to fully elute Triton X-100, then perfuse with 1% SDS perfusion solution at a rate of 10ml / min for 3 hours, at this time the liver becomes transparent, and the vascular structure in it can be ...
Embodiment 2
[0032] Example 2. Pig liver perfusion decellularization
[0033] 1. Prepare perfusate I and perfusate II:
[0034] Perfusate I: Dissolve 100ml of Triton X-100 in 800ml of Tris-HCl with pH 8 and 50mM, and dilute to 1000ml to obtain 10% Triton X-100;
[0035] Perfusate II: Dissolve 100 g of SDS in 800 ml of Tris-HCl at pH 8 and 50 mM, and dilute to 1000 ml to obtain 10% SDS.
[0036] 2. Perfusion to achieve decellularization:
[0037] After the portal vein and the hepatic artery were respectively intubated, the above-mentioned perfusate I and perfusate II were perfused simultaneously until the liver became transparent. The perfusion rate used was 50 ml / min. After perfusion, perfuse with double distilled water to fully elute the detergent.
Embodiment 3
[0038] Example 3. Mouse liver decellularization
[0039] 1. Prepare perfusate I and perfusate II:
[0040] Perfusate I: Dissolve 1ml of Triton X-100 in 800ml of Tris-HCl with pH 7.5 and 50mM, and dilute to 1000ml to obtain 0.1% Triton X-100;
[0041] Perfusion solution II: 1 g of ursodeoxycholic acid was dissolved in 800 ml of Tris-HCl of pH 7.5 and 50 mM, and the volume was adjusted to 1000 ml to obtain 0.1% ursodeoxycholic acid.
[0042] 2. Perfusion to achieve decellularization:
[0043]After passing through the superior and inferior hepatic vena cava, the inferior hepatic inferior vena cava was ligated, and the portal vein was cut to perfuse at a rate of 4ml / min. Perfuse according to the method in Example 1.
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