Microfluidic system for simulating lung tissue
A microfluidic system and lung-simulating technology, applied in tissue culture, tissue cell/virus culture devices, bone/connective tissue cells, etc., can solve problems such as cell death, and achieve the effect of in vitro diagnosis and customized drug prescription
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Embodiment 1
[0061] Example 1 Isolation of lung epithelial cells and lung fibroblasts from human lung tissue
[0062] To fabricate a microfluidic system that mimics in vivo lung tissue, lung epithelial cells and lung fibroblasts were isolated from human lung tissue by the following method.
Embodiment 1-1
[0063] Example 1-1 Dissociation of human lung tissue
[0064] 10 normal lung tissue samples (N-1 to N-10) of 0.39 g to 1.37 g in Table 1 below obtained from normal lung tissue in resected lung tissue of lung cancer patients who had undergone surgery, and from lung cancer patients 2 lung cancer tissue samples (C-1 and C-2) with a total of 0.41 g in Table 1 below obtained from cancer tissue in resected lung tissue were stored in storage buffer (MiltenyiBiotec, Tissue Storage Solution) at 4°C for 1 From day to day 2, these tissue samples were then divided into 0.4g to 0.5g each and put into each C tube (C test tube) respectively.
[0065] 【Table 1】
[0066] tissue sample number Weight (g) tissue sample number Weight (g) N-1 0.54 N-7 1.37 N-2 0.52 N-8 0.64 N-3 0.66 N-9 1.00 N-4 0.70 N-10 0.39 N-5 1.02 C-1 *
0.28 N-6 1.17 C-2 *
0.13
[0067] *The total weight of lung cancer tissue samples C-1 and C-2 was 0.41 g, ...
Embodiment 1-2
[0071] Example 1-2 Isolation of lung epithelial cells and lung fibroblasts
[0072] Human lung tissue dissociated in Example 1-1 Lung epithelial cells and lung fibroblasts were isolated by the procedure described below.
[0073] Specifically, calculate the number of cells contained in the T-175 flask in Example 1-1, prepare 5X10 6 to 1X10 7The cells were centrifuged at 300×g for 10 minutes at room temperature to prepare three suspensions of cell clumps, ie, a total of 300ul of lung epithelial cell suspension and a total of 80ul of lung fibroblast suspension. At this time, the autoMACS Rinsing solution (Miltenyi Biotec, #130-091-222) and the MACS BSA stock (stock) solution (Miltenyi Biotec, #130-091-376) were combined at a volume ratio of 20:1, that is, using the The three suspensions were prepared with PEB buffer obtained by mixing 47.5 ml of autoMACS wash solution and 2.5 ml of MACS BSA stock solution.
[0074] Then, in order to improve the purity of the isolated cells, 10...
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