Room Carry Interleukin 18 Gene In Human Umbilical Cord Mesenchymal Stem Cells And Its Significance

Objective To construct IL-18gene in human umbilical cord mesenchymal stem cells for exploring a means of tumor targeted gene therapy research.Methods We isolated and cultured hUMSCs in vitro. The immunophenotype of hUMSCs like CD29.CD44, CD105,CD34and CD45was detected by flow eytometry. The lentivirus vector containing human IL-18gene and the blank control lentivirus were constructed and transfected into hUMSCs. There is a total of three groups:the experimental group transfected with a lentivirus (IL-18-hUMSCs), transfected with blank control group lentivirus (NC-hUMSCs) and the control group did not transfected (hUMSCs). The IL-18mRNA and protein expression level of every group was detected by semi-quantitative RT-PCR and Western blot.Results We isolated and cultured hUMSCs successfully. The FACS indicated that CD29.CD44and CD105were positive; however the CD34and CD45were negative. So it med the phenotype of hUMSCs. IL-18-hUMSCs was produced successfully. Comparing with other groups, the expressions of IL-18protein and mRNA in experimental group were significantly increased (P<0.05). RT-PCR and Western blot showed that IL-18gene was successfully transfected into hUMSCs and stable expressed.Conclusion Lentivirus-IL-18vector can be successfully transfected and stablely expressed in hUMSCs that it provided a new experimental method of tumor targeted gene therapy experimental study.