Gene Expression Profile of Immunoregulatory Cytokines Secreted by Mesenchymal Stem Cells Obtained from Different Tissues

Background and Aim: Mesenchymal stem cells have a long-term clinicalapplication and are well-known due to their immunomodulatorycharacteristics. Many clinical applications have been proposed in arraysof studies, therefore, we attempt to investigate which mesenchymal stemcells source and passage can provide the best possible immunoregulatoryresponses in patients.Methods: We gathered samples of adipose tissue and bone marrow fromvolunteer donors. Bone marrow mononuclear cells were isolated byFicoll, the cultivation of cells carried out standard conditions (5% CO2,37°C). MSCs isolated were identified according to phenotype, physicalproperties, and cell surface markers. MSCs were attached to the plasticsurface of the flask and getting fibroblast shapes. To characterize themas a Mesenchymal stem cell CD73, CD90 and CD105 as appositive markers and CD14, CD34 and CD45 were considered by flow cytometry.Four different passages of stem cells contain 3, 5, 7 and 9 selected asdifferent groups to study the expression level of cytokines. All wereutilized for RNA extraction and cDNA synthesis. Expression of themain immunomodulatory cytokines ICAM, IDO, VCAM, TGF-beta, IL-6,IL-10, PGE2 was measured by real-time PCR. Expression levels of allcytokines were analyzed according to comparative analysis method byRest software.Results: Totally MSCs were isolated from 8 donors. They were equallyobtained from bone marrow and adipose tissue. Expression level Variationof cytokines in different passages was observed. In addition, cytokinepanel of Mesenchymal stem cells with different source was not identical.According to the bone marrow-derived MSCs Real-Time PCR results;IDO, VCAM expression in passage 3, PGE2, ICAM, IL-6 gene expressionin passage 5, and TGF-beta in passage 9 were more pronounced. Onthe other hand, in adipose tissue sources, the expression of ICAM, IDO,VCAM, IL-6 genes in passage 7 was up-regulated.Conclusion: MSCs are the reliable source of stem cells in clinicalapplication. However, some MSCs derived cytokines stimulate diseaseprogression. IL-6 could be nominated as a stimulating factor in LiverFibrogenesis. So, investigating in the immunoregulatory mechanism ofMSCs is demanded. According to our results, passage 7 in Adipose tissueMSCs and passage 3 and 5 of the Bone marrow MSCs would be betterin clinical applications of autoimmune disorders. In other side Passagenumber 5 of BM-MSCs seems not to be a suitable candidate for fibroticdiseases.

Keywords:

MSC , Immunoregulatory , Adipose tissue , Bone marrow , Cytokine