Method for culturing contrast agent damage model based on renal tubular epithelial cells

Abstract

The invention discloses a method for culturing a contrast agent damage model based on renal tubular epithelial cells. The method includes the steps of: (1) conducting HK-2 cell resuscitation; (2) performing cell subculture; (3) observing the culture period cell morphology under a microscope, when the cell edge crumples become round and the refractivity is enhanced, conducting centrifugation, and adding a complete medium to make a cell resuspension solution; (4) transferring the cell resuspension solution into a complete medium according to an inoculation density of 40%-50%; (5) when the cell growth density reaches 80%-90%, performing centrifugation, adding a complete medium to prepare a cell resuspension solution (with a cell concentration of 1.0-1.5*10<6> / ml); and (6) inoculating every 40-60microl of the prepared cell resuspension solution into a 2ml complete medium, conducting culture for 40-50h, then conducting replacing with a serum-free medium and performing culture for 4-8h, thenadding a 45-55mgI / ml iodixanol solution, and further conducting culture for 5-7h, thus obtaining damage model cells. The method provided by the invention can culture the cell damage model that has similar content to contrast-induced nephropathy patient postoperative serum HMGB-1 content, closer damage and fewer system toxins.

Description

technical field [0001] The invention relates to the technical field of contrast agent cell injury model preparation, in particular to a method for culturing a contrast agent injury model based on renal tubular epithelial cells. Background technique [0002] Due to the irreplaceable advantages of angiography and interventional diagnosis and treatment techniques in disease diagnosis, the use of iodine contrast agents is becoming more and more widespread. However, the incidence of contrast-induced nephropathy is on the rise. So far, contrast-induced nephropathy has become the third leading cause of iatrogenic renal failure, second only to renal insufficiency and nephrotoxic drugs in incidence. [0003] The mechanism of contrast-induced nephropathy is not completely clear, among which serum high mobility group box-1 (HMGB-1) is a highly conserved nuclear protein discovered in recent years, its main function is to stabilize the chromosome structure and assist its function, and p...

AI Technical Summary

Problems solved by technology

Due to ethical issues, in-depth research cannot be carried out directly in the human body, and the molecular mechanism often needs to be confirmed by in vitro cell experiments. However, although there have been reports of human renal tubular epithelial cell injury by adding contrast agents to in vitro cell culture fluid, the obtained It’s just that the severely damaged human renal tubular epithelial cells are very different from the damaged cells and extracellular environment in patients with contrast-induced nephropathy, which is not conducive to the development and testing of related drugs, and even leads to errors in the results of related drug tests

Therefore, no stable research model has been found to establish a suitable HMGB-1-induced injury to renal tubular epithelial cells