Construction Of Two Drugs Three Concentration Gradient Microfluidics And Its Application In Tumor Cells Analysis

Cancer,malignant tumor,contains hundreds of related conditions.It is a "killer" status for human health due to low cure rate and high fatality rate for humanity.Treating cancer is a long way for humans Chemotherapy poses a great danger to humans as using highly toxic drugs.With the utilization and development of traditional Chinese medicine resources,a growing number of drugs have been researched because of low-toxic and anti-cancer effects.However,traditional method for cancer cells requires a large amount of reagents and consumables.It is time-consuming and laborious.Therefore,emerging microfluidic chips have developed rapidly in recent years.In this study,a new type of microfluidic chip was used to conduct a series of experiments on the concentration gradient formed.Then,two cancer cells(human breast cancer MCF-7 cells and human hepatocarcinoma HepG2 cells)and two anti-cancer drugs(Doxorubicin and Cisplatin)were applied to the microfluidic chip.Whereafter,traditional methods were used to study the effects of two medicinal plants(Taraxacum mongolicum Hand.-Mazz.and Viola yedoensis Makino)on the above two tumor cells.Moreover,we analyzed the cell proliferation and apoptosis by various identification methods.Finally,we compared the results of microfluidic chip method and the traditional method.The main findings were presented as follows1.A single-layer microfluidic chip was designed and produced.We conducted quantitative analysis of the concentration gradient with different fluorescent dyes and digital software.It was proved that the chip formed 12 concentration gradients which were similar to the digital simulation results.The chip achieved the required concentration under a wide range of Rec(0.02-40.53).Stability of the chip could be maintained for more than 5 days.Its ability to research cells was proved by fluorescence intensity verification of FN modification,the study of flow rate and mixing effect and the concentration stability analysis.2.Both cancers cells(MCF-7 and HepG2)can grow normally and have good morphology.in the microchambers of the microchip.Using AO/PI staining method,we identified the vitality of the two cells in the chip.The significance of cell growth and apoptosis were researched under conditions of continuous perfusion of conventional DMEM media and drug media.It was demonstrated that the dependence of the two tumor cells on the time and concentration of anticancer drugs.And it was verified that the ability of cell sorting,drug screening and explore the drug ratio.3.We investigated the viability of MCF-7 and HepG2 cells(traditional cell culture methods)in response to two antitumor medicinal plants(Taraxacum mongolicum Hand.-Mazz.and Viola yedoensis Makino).AO/PI double staining,MTT assay,and mitochondrial membrane potential assay were used to research inhibition of proliferation and mitochondrial membrane potential changes.After 48 hours of the effects above two Chinese herbal extracts,the activity of both tumor cells was reduced to about 6%to 7%and the inhibition rate was over 92%.Subsequently,the combination of two drugs,microfluidic chip and AO/PI staining method proved that the chip could be used for cell sorting,drug screening,drug lethality,etc..In fact,it had huge advantages over traditional methods in economic and time.In summary,this study provides a new idea for microfluidic chip miniaturization,practicality and versatility.It provides a new method for researching anti-tumor drugs.Moreover,it has more efficient and economical effect on screening drug and sorting cells.