High-Precision Detection Of Cell Cross-Contamination And Its Functional Study

Organs,tissues or cells taken from the body simulate the physiological environment of the body in vitro,that is,under the conditions of suitable p H,osmotic pressure,temperature,sufficient nutrients,etc.,the process by which cells can survive,grow and reproduce is called in vitro Cell culture,also known as cell cloning technology.Since the in vitro cell culture technology was invented,more and more biological researches are inseparable from cell culture,and with the development of virology,immunology,cell biology,molecular biology,genetics,toxicology and pharmacology,etc.The establishment of disciplines,cell culture technology is becoming more and more important,and it is not an exaggeration to regard cell culture technology as the cornerstone of biology.However,since Hela cells were discovered and successfully cultured,the pollution problem of the cell culture process has also arisen,especially the cross-contamination problem between cells,which has caused great troubles for scientific researchers.Although the method of identifying cell cross contamination appeared as early as the 1960 s,there are still many problems.The methods to identify cell cross-contamination mainly include isozyme mapping method,human leukocyte antigen(HLA)typing method,short tandem repeat sequence(STR)mapping method,etc.,but the detection accuracy is worrying.According to reports,when cells cross When the pollution is less than 10%,none of these methods can detect it.Crosscontamination of cells below 10% is called light contamination.This experiment is to process the sequencing data after simulating cell cross-contamination,STR site amplification,database construction and sequencing,and sort it into the sequencing data processing program developed by our laboratory,and we can get that the experimental group has cell cross-contamination.It can prove that this method of identifying cell cross-contamination is feasible,and its detection accuracy can reach 0.5%,that is,contaminated cells account for 0.5% of the total number of cultured cells.By simulating the slight cross-contamination of cells,the impact on cell function experiments is discussed,mainly the impact on cell proliferation.It is found that in the case of 5% cell cross-contamination,cell cross-contamination will affect the proliferation rate of cells.It may have a small impact in the first few days of cell culture.Once the contaminated cells begin to multiply,it will affect the original cells.Proliferation has a huge impact,and may even cause the death of original cells.The consequences of cell cross-contamination should be taken seriously by various scientific research units.