Artificial pancreas islet tissue and preparation and application thereof
A pancreatic islet, artificial technology, applied in tissue culture, biochemical equipment and methods, pancreatic cells, etc., can solve the problem of inevitable drug dependence, drug toxicity and side effects, etc.
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Embodiment 1
[0062] Embodiment 1: the configuration of mixed culture medium
[0063] The composition of the mixed culture medium is: RPMI1640 + 10% FBS + 1% penicillin + 1% streptomycin, the culture medium is placed at 37°C, 5% CO 2 Equilibrate in the incubator for 2-4 hours.
Embodiment 2
[0064] Example 2: Obtaining pancreatic islet cells
[0065] The mice were sacrificed by neck dislocation, the entrance of the common bile duct to the duodenum was ligated, and the common bile duct was perfused with collagenase P 2.4 mg / mouse. After the pancreas was filled, the whole pancreas was carefully taken out; Shake and digest at 37°C for 12 minutes; add pre-cooled 10% FBS RPMI1640 medium after digestion to terminate digestion, shake vigorously for 15 seconds, then centrifuge at 4°C and 800 rpm for 4 minutes to remove supernatant; resuspend sediment in 10% FBS RPMI1640 medium and filter through a 50-mesh metal sieve , the filtrate was centrifuged at 800rpm at 4°C for 4min to remove the supernatant; resuspend the pellet in 10ml histopaque 1077 and slowly add 10ml serum-free RPMI1640 medium to make a clear layered interface between the two solutions; centrifuge at 2400rpm at 20°C for 20 minutes, and absorb the tissue between the two layers For the cell mass, wash 1-2 times...
Embodiment 3
[0066] Example 3: Preparation of 3D printed islet tissue
[0067] 1. Design and preparation of biological scaffolds
[0068] (1) Combining with computer aided design (CAD), three-dimensional printing technology is used to construct the bracket, in which the shape, composition and internal structure are well designable;
[0069] (2) The shape of the three-dimensional islet tissue is designed to be similar to that of the pancreas, and the internal structure is guaranteed to have a certain pore structure;
[0070] (3) The scaffold material is printed with gelatin compatibility.
[0071] (4) Internal structural parameters of the constructed three-dimensional islet tissue: pore size (R): = 0.2mm; line stacking angle: 90°.
[0072] (5) Store the prepared scaffold at -80°C for future use.
[0073] 2. Preparation of 3D artificial islet tissue
[0074] Each mouse pancreas contains 600-4000 islet tissues, and each islet tissue contains about 1000 islet cells. During printing, a thre...
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