Kit for screening drug developmental toxicity based on C/EBP[alpha] and IGF1R gene and application thereof
A technology for screening drugs and kits, applied in the field of drug toxicology, can solve the problems of unclear developmental toxicity of drugs, lack of theoretical system of mechanism of occurrence, etc., achieve high sensitivity, fast detection process, and wide application range
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Embodiment 1
[0050] The effect of a kit for detecting drug developmental toxicity based on C / EBPa and IGF1R genes on the developmental toxicity drug methimazole in the L02 cell line model.
[0051] The kit consists of detecting intracellular C / EBPa and IGF1R gene expression, and simultaneously detecting the luciferase activity of the IGFIR promoter and luciferase reporter gene. The above-mentioned kit for detecting drug developmental toxicity based on C / EBPa and IGF1R genes is applied to the developmental toxicity drug methimazole in the L02 cell line model as follows:
[0052] 1 Preparation of human fetal L02 cells
[0053] 1.1 Take out the cell cryopreservation tube from the liquid nitrogen tank, put it into a 37°C water bath quickly, and shake it from time to time to thaw as soon as possible.
[0054]1.2 After complete thawing, centrifuge at 1300rpm for 3min, wipe and disinfect the cryopreservation tube with 75% alcohol, and then move it to a biological safety cabinet.
[0055] 1.3 As...
Embodiment 2
[0094] The effect of a kit for detecting drug developmental toxicity based on C / EBPa and IGF1R genes on the developmental toxicity drug methimazole in WJ-MSCs hepatic-like differentiation cell model.
[0095] 1. WJ-MSCs primary extraction and hepatic differentiation
[0096] 1.1 WJ-MSCs primary extraction
[0097] Under aseptic conditions in the operating room, the umbilical cord of the fetus delivered by cesarean section was taken out, the blood was washed, soaked in pre-cooled saline, and brought back to the laboratory. The umbilical cord was taken out in the ultra-clean workbench, and all separation processes were performed in pre-cooled saline. Cut into 4-6cm / section, and wash away the blood with a syringe. Remove the umbilical cord adventitia, umbilical artery, and umbilical vein, and peel off Wharton's glue: first, fix the two ends of the umbilical cord on a wooden board with tacks, and use ophthalmic scissors to separate the adventitia longitudinally from one end, and...
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