The Study Of The Isolation, Culture And Identification Of MSCs Of Psoriatic Patients Lesions And Its Effect On T Cell Proliferation

[Objective]:To establish skin MSCs (mesenchymal stem cells) culture system through MSCs of psoriatic patients isolated, cultivated and identified in vitro, and to study the impact of MSCs that in psoriatic patients on peripheral blood T cell proliferation,and the relation between MSCs cytokine levels of psoriatic patients skin and the degree of psoriatic.[Methods]:①The mononuclear cells were isolated in15psoriatic patients lesions and in15healthy controls skin respectively.②The cell surface markers of CD29, CD34、CD44、CD45、CD73、CD90、CD105and HLA-DR were detected by flow cytometry.③The multi-directional differentiation cells were identified by induced cells through corresponding inducers.④The effects of MSCs on peripheral blood T cell proliferation was observed by MTT.⑤The cytokine levels in third-generation skin a single nuclear cell culture supernatant were detected by ELISA.⑥According to Psoriasis Area and Severity Index (PASI) of psoriatic lesion area and extent, the correlation of cytokine levels and psoriatic lesion degree was assessed by the Pearson correlation analysis.[Results]:①MSCs were isolated and cultured:a. divided skin organization into small pieces, placed in the2.5g/L Dispase II enzymes,37℃for three hours, separated dermis and epidermis, abandoned the epidermis, the dermis chopped, percussion, filtration, centrifugation, collection and cultivation in37℃and5%CO2incubator; B. spread to the third generation when cell wall of more than90%, C. Two groups of MSCs were observed under inverted microscope.②Flow cytometry showed that the MSCs surface markers CD29、CD44、CD73、CD90and CD105were highly expressed, and CD34、CD45and HLA-DR were expressed lower, which proved that cultivate MSCs have higher purity.③MSCs can be differentiated to adipose cell,osteoblast and chondrocytes via induction.④T cell proliferation activity test results:MSCs in psoriatic patients lesions and normal skin has inhibitory effect on peripheral blood T cell proliferation (P<0.05), but the inhibition weakened in psoriatic patients lesions (P<0.05).⑤The level of IL-11in psoriatic patients MSCs supernatant fluid was significantly higher than that in the normal control group (P<0.05).While the levels of IL-6and HGF was significantly lower compared with normal control group (P<0.05).⑥There is no correlation between MSCs cytokine levels in psoriatic patients skin and PASI score.[Conclusion]:①isolated, cultured and identified MSCs of skin, Establish the cultivation system of skin MSCs, provide correct and effective method for training skin MSCs.(2)MSCs derived from psoriatic skin lesions had weaker inhibitory effect on T cell proliferation, which may be indirectly promote the psoriatic skin occurrence and development of the inflammatory response in the micro environment.③The decreased inhibition of MSCs on peripheral blood T cell proliferation may be due to the abnormal secretion of cytokines, and cytokine levels no correlated with PASI score. This revealed abnormal MSCs in psoriatic patients may be involved in the pathogenesis of psoriasis.