Effect Of Human Umbilical Cord-derived Mesenchymal Stem Cells And Oxaliplatin On The Growth Of HepG2Cell Lines

Objective:Using Transwell co-culture techniques to study the effect of hUCMSCs and oxaliplatin on thegrowth of HepG2cells, to explore hUCMSCs and oxaliplatin whether can promote apoptosis ofHepG2cells together.Methods:Collagenase II digested umbilical cord tissue to screened hUCMSCs. We can identifyhUCMSCs by observing its morphology, detecting the cells’growth curve and identifying itsphenotype. To detect proliferation inhibition rate of HepG2caused by oxaliplatin by cell countingkit-8(CCK-8)assay. Transwell culture experiments were divided into four groups, the first group:hUCMSCs were inoculated in the upper chamber and HepG2cells were seeded in the lowerchamber; the second group: the upper and lower chambers were inoculated hUCMSCs and HepG2cells respectively, adding oxaliplatin with a fixed concentration(20ug/ml) in the lower chambers;the third group: HepG2cells were seeded lower chamber with oxaliplatin (20ug/ml), adding thesame amount of DMEM complete medium; the fourth group: HepG2cells were seeded lowerchamber with the same amount of DMEM complete medium in the upper chamber. To inoculatelogarithmic phase of hUCMSCs and HepG2cells by above grouping and to detect HepG2cells’apoptosis rate on48h or72h later by flow cytometry.Results:The cells separated from umbilical cord by collagenase II were homogeneous fibroblast-likecells in swirling or radiation growth after passaging. The detection of cell growth curve shows:After the first two days, passaging,cells were in lag phase, In the third to fifth days, cells were inlogarithmic phase, When time went to the sixth to seventh days, cells were in plateau period. Thephenotype analysis of cells separeted from umbilical cord shows they are positive for markers suchas CD44、CD73、CD90、CD105; at the same time, they express negligible levels of CD31、CD34、CD45、HLA-DR. The data from CCK-8assay measured by general linear model-repeated measures shows thattime effect has statistically significant(P<0.05); the interaction effect of time and concentration alsohas statistically significan(P<0.05); different drug concentration groups have the differentproliferation inhibition rate (P<0.05).The data from Annexin V-FITC/PI measured by general linear model-repeated measures andANOVA two-way factorial design shows that time effect has statistically significant(P<0.05); theinteraction effect of time and grouping also has statistically significant(P<0.05); different groupingshave different apoptosis rate(P<0.05); there is interaction between hUCMSCs and oxaliplatin on theinduction of apoptosis in HepG2cells in the two periods(P<0.05).Conclusions:1. Cells separated by Collagenase II have the same morphology, growth and phenotype withhUCMSCs, So we can purify the hUCMSCs by Collagenase II.2. HepG2cells’proliferation can be restrained by oxaliplatin in a time-dependant anddose-dependant.3. The results reveals that oxaliplatin can promote apoptosis of HepG2cells and showed atime-dependent manner, hUCMSCs cells can promote apoptosis of HepG2cells and showed atime-dependent manner, hUCMSCs and oxaliplatin can enhance effectors of promoting apoptosis ofHepG2cells together.