Preliminary Study On The Differentiation Of Bone Marrow Mesenchymal Stem Cells In Psoriatic Patients To Vascular Endothelial Cells In Vitro

【Objective】In our study, the activity of psoriatic blood vessel endothelial cells (VEC), oriented differentiated from patients bone marrow mesenchymal stem cells (BMMSCs) in vitro,was detected in purpose of providing some basis for the further researches of BMMSCs biological characteristics in patients with psoriasis.【Methods】①BMMSCs of patients with psoriasis and control were gained after density gradient isolation combined with adherence isolation. The phenotypes of the cells were identified by flow cytometry .②To oriented differentiate the BMMSCs into VEC, Vascular endothelial growth factor(VEGF) and basic fibroblast growth factor (bFGF) was added into endothelial cell growth medium (DMEM). The induced BMMSCs were detected by flow cytometry to find whether they had endothelial cell phenotypes.③The expression of endothelial-specific markers such as VWF and KDR were assayed by immunofluorescence cytochemistry④The characteristic capillary-like structures forming ability of differentiated cells was tested using in vitro angiogenesis test kit.⑤Transwell chamber in vitro migration assay was used to detect migration capacity of induced VEC to T cell.⑥A s in one patient whose parents were both patients with psoriasis, the BMMSCs were found to differentiate into VEC in the primary culture process. High-throughput RNA sequencing technologies was taken advantage of to screen the differentially expressed genes (DEGs).【Results】①Flow cytometry results showed that surface antigens CD44 and CD29 were highly expressed while CD34,CD45,HLA-DR were negatively expressed in the BMMSCs, which suggested that the cultured BMMSCs were of high purity.②The expressions of CD34, HLA-DR ,CD54 and CD45 were weak positive in both test group and control. And the expression level of test group was significantly higher than control (P<0.05).③Immunocytochemistry results showed that both the two groups can differentiate into vascular endothelial cells.④The capillaries forming ability in semisolid medium in patients was higher than in control⑤The number of T cell migration from patient with psoriasis are larger than that from normal contrasts.⑥A total of 475 DEGs were identified in from this patient, of which 298 were up-regulated and 177 down-regulated. Gene ontology analysis showed that 2 gene terms involving in prostanoid metabolic process and prostaglandin metabolic process were significantly enriched (P<0.05). 【Conclusion】The psoriatic BMMSCs biological characteristics were abnormal and they were prone to differentiate into VEC. BMMSCs play roles in the attack of psoriasis, and their ability that can differentiate into VEC abnormally may contribute to the abnormal growth of microvascular proliferation of papillary layer in lesional dermis in patients.