The Study Of Inducing Human Adipose Derived Mesenchymal Stem Cell-hematopoietic Stem Cells Differentiation To CIK Cells

Objects:(1) This project aimed to build a modified method to isolate and culture thehuman adipose-derived mesenchymal stem cells and umbilical cord mesenchymalstem cells and to compare the growth properties between these two kinds ofmesenchymal stem cells.(2) This project explored the potential of the differentiation ability from theAdipose Derived Mesenchymal Stem Cell-Hematopoietic Stem Cells（ADMSC-HSCs）to CD3+CD56+lymphocyte, which is also named as the cytokine-inducedkiller cells(CIK cells), under the stimulation of multiple cytokines.Methods:(1) The primary mesenchymal stem cells were isolated and pured from thehuman donor’s adipose and umbilical cord samples by the modified methods andadherent culture. Then we analyzed the characteristics of the primary and passageADMSCs and UCMSCs:The cell morphological differences were observed and compared throughinverted microscope and Giemsa’s staining. The MTT assay was used to draw thegrowth curve and compare the growth rate. And the flow cytometry was used to testthe surface markers of mesenchymal stem cells from two different origins.(2) The third passage ADMSCs were induced to re-differentiate into HSCsthrough specific medium after transfected by the shRNA-337.5days later, theexpressions of CD34, CD38, CD45which are the cell surface marker of HSCs, werequantified by flow cytometry. Then the ADMSC-HSCs were harvested and changedthe culture environment to the CIK-induced medium. After7or14days induction,the tests of the CIK cell characteristic surface marker CD3and CD56wereperformed.Results:(1) The ADMSCs and UCMSCs could be successfully acquired using themodified methods. The primary ADMSCs were easier to acquire than the UCMSCs. Both kinds of mesenchymal stem cells had the typical short shuttle-like and arrangedin swirl with adherence. The UCMSCs were bigger and growing faster than theADMSCs. Both ADMSCs and UCMSCs highly expressed CD29, CD44, CD90,CD166, but no CD34, CD45, CD49f and CD133.(2) The ADMSCs’ morphology and properties were both changed aftertransfected with shRNA-337and re-differentiate inducing cultured5days. TheADMSC-HSCs became colony-like suspending in the medium. The flow cytometryassay demonstrated that the ADMSC-HSCs could express CD34, CD38, CD45, butno CD29and CD166. Then, after the CIK-induced culture for14-days, theADMSC-HSCs became to express the CIK cell characteristic surface marker CD3and CD56. The CD3+CD56+rate can reach (25.12±1.38)%.Conclusion:(1)The modified methods to isolate the ADMSCs and UCMSCs are established.Both the morphology and properties have parts of differences between these twokinds of mesenchymal stem cells. The growth rate of UCMSCs is faster thanADMSCs. And these two kinds of mesenchymal stem cells share the same stem cellcharacteristic surface makers.(2) The ADMSC-HSCs have the capability to differentiate into cytokine-inducedkiller cells.