| In cell biology,the main methods for detecting cell activity and proliferation are the 3 H-TdR incorporation and MTT colorimetry method.3 H-TdR incorporation method is a method that directly through dividing cell numbers to evaluate cell proliferation.MTT method is an indirect method,namely through the detection of cell viability according to the number of healthy cells in test samples,and then evaluate the ability of cell proliferation.MTT is a frequently-used method for detecting cell survival and growth.The principle for MTT assay is that Succinate dehydrogenase in mitochondria of living cell can make the exogenous MTT be reduced to water insoluble purple crystal deposition called formazan in cells,however,dead cells can not produce the succinate dehydrogenase.By enzyme-linked immunosorbent assay to detect the absorbance in 492 nm wavelength,we can construct a relationship between the number of formazan formation and the living cell number,so as to obtain the cell viability.Researches have showed that in a certain concentration range of cells,the absorbance is proportional to the number of living cells.This method has been widely used in detecting the activity of some bioactive factors,mass screening the anti-tumor drugs,and the cell toxicity test.MTS is a new type of four azole compounds.The principle of MTS is the same as MTT,but its formazan is water-soluble,so the experimental operation don't need to dissolve formazan by DMSO like the MTT method,so it is much more efficient compared with the MTT method.In addition,the color of MTS products is deeper,the absorbance variation range is larger,and it has higher sensitivity and more accurate results.MTS method could more quickly and accurately determine cell proliferation,and also avoid errors caused by manual operation or test environment,it greatly improves the accuracy of the test results.This paper is divided into two parts,firstly,we use the MTT method and cell imaging technology to study the bio-compatibility of inorganic nanomaterials-fHMSN,then we use the anticancer drug doxorubicin(DOX)as a drug model,to study how can the fHMSN take the drug into cells and release the drug gradually.In addition,we also use the MTS method and cell imaging technology to research the influence of small molecular organic carbon solution(AC)on proliferation of in-vitro cultured cancer cells and human placenta mesenchymal stem cells(PMSC).The first part of the results show that the hollow mesoporous silica nanospheres fHMSN at a concentration of less than 50ug/ml,the cell viability is >90%,so the fHMSN has low toxicity,and can be used as a carrier of anticancer drugs in clinic.After fHMSN loading the anticancer drug doxorubicin into cancer cells,it can gradually release the drug,to avoid toxic side effects of high doses of the drug.Then using cell imaging technology,it can be observed that with the incubation time goes on,fHMSN can be absorbed into the cells by endocytosis,the drug loaded by fHMSN can also be gradually endocytosed into cells,and mainly resists in the cytoplasm of cells.In the second part of the experiment,we use MTS method to confirm that the anti-tumor effect of small molecule available carbon(AC)is not through direct effection on tumor cells.The experiments also found that although the small molecule available carbon(AC)has no significant inhibitory effect on tumor cells,it can promote normal cell proliferation in vitro,so we select human mesenchymal stem cells whose growth rate was very slow when cultured in-vitro under normal culture condition,after add different dilutions of AC into the medium,the experimental group of mesenchymal stem cell's growth is good,and the growth rate has accelerated noticeably,compared with the control group.This finding is expected to solve the slow proliferation rate of mesenchymal stem cells(MSCs)when cultured in vitro,it can provide a possible solution for the efficient in vitro culture of mesenchymal stem cells(MSCs). |
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