Non-alcoholic fatty liver disease cell model induction liquid and preparation method and application thereof
A fatty liver disease, cell model technology, applied in biochemical equipment and methods, animal cells, tumor/cancer cells, etc., can solve the problems of high requirements, complex methods, long modeling cycle, etc., to achieve a high modeling success rate Effect
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[0020] The invention provides a method for preparing a non-alcoholic fatty liver disease cell model inducing solution, which comprises the following steps:
[0021] 1) After mixing stearic acid and sodium hydroxide solution, saponification is performed to obtain a saponification liquid; the mass-volume ratio of the stearic acid and sodium hydroxide solution is 1g: (170~185) mL;
[0022] 2) After mixing the saponification solution and the fat-free bovine serum albumin solution with a mass concentration of 35-43% in equal volume, the induction solution mother liquid is obtained;
[0023] 3) Dilute the mother liquor of the induction liquid with a fetal bovine serum medium with a volume fraction of 7-12% to obtain an induction liquid.
[0024] In the preparation method of the non-alcoholic fatty liver disease cell model induction solution of the present invention, stearic acid and sodium hydroxide solution are mixed and then saponified to obtain a saponified solution; the mass volume of th...
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[0029] Example 1
[0030] 1) Prepare 40% fat-free bovine serum albumin: preheat the PBS solution to 55°C, measure 1.2g of fat-free bovine serum albumin, place in a 15mL centrifuge tube, add 2mL PBS solution, do not shake or mix, just put it directly Centrifuge in a high-speed centrifuge at 8000 rpm for 20 minutes to completely dissolve, and dilute to 3 mL with PBS. Obtained 3 mL of fat-free bovine serum albumin solution with a mass concentration of 40%, which was brown and yellow in appearance;
[0031] 2) Prepare 20mM stearic acid solution: take 0.04g sodium hydroxide, dissolve it in 10mL deionized water, make 10mL of 0.1mol / L sodium hydroxide solution, take 3mL. Weigh 0.017 g of stearic acid into 3 mL of sodium hydroxide solution, place it in a water bath at 100°C for full saponification for 10 minutes, and finally become colorless, transparent and clear to obtain 20 mM sodium stearate saponification solution.
[0032] 3) The warmed stearic acid solution is quickly added to the s...
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[0033] Example 2
[0034] Dilute the 10mM stearic acid solution 50, 100, 200 times with 10% fetal bovine serum medium to form stearic acid working solutions with concentrations of 200μM, 100μM, and 50μM, respectively, for growth in logarithmic phase HepG2 cells were treated for 24h, the above experiment was repeated 3 times, stained with oil red O, the results are as follows figure 1 As shown, it can be seen that 200 μM, 100 μM, and 50 μM stearic acid working solutions can all be successfully modeled.
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