The Effect Of Decitabine On Megakaryocyte Culture Of Steroid-resistant ITP Patients

Objective:In this study, we investigated the effect of decitabine on megakaryocyte cultrue in an in vitro megakaryocyte differentiation model utilising human bone marrow cells of Steroid-resistant ITP patients, and the effect of ITP plasma on megakaryocyte maturation was observed in the same way.Materials and methods:20cases of Steroid-resistant ITP patients were inpatients and outpatients of Zhuzhou Central Hospital from March2013to April2014. Both methyl cellulose semisolid culture system (to observe and count the number of megakaryocytes colony-forming unit) and liquid culture system (to analysis the expression rate of CD41a+cells) were used. The experiment was divided to four groups according to the different components of the culture system, Group A was control, Group B added decitabine, Group C added ITP plasma, Group D added both decitabine and ITP plasma, and the rest of the culture components were the same in the four groups except above. Morphous of megakaryocytes was observed with an inverted microscope and light microscope.The expression rate of CD41a+cells in culture was analysed by flow cytometric.Results:(1) Different concentrations of decitabine had different effect on megakaryocyte growth of Steroid-resistant ITP patients and the best concentration to differentiate bone marrow mononuclear cells into megakaryocyte is3.0uM.(2) Compared with Group A, both megakaryocyte colony and expression rate of CD41a+cell were higher in Group B, P<0.05, the difference was statistically significant.(3)The megakaryocyte colony was lower in Group C than Group A, the difference was statistically significant;(4)Megakaryocyte colony was higher in Group D than Group C;(5) There was no statistically significant difference between Group B and Group D.Conclusions:(1) Decitabine is able to induce bone marrow mononuclear cells of Steroid-resistant ITP patients to differentiate into megakaryocyte and the best concentration is3.0uM (2) ITP plasma is able to inhibit the growth of megakaryocyte of Steroid-resistant ITP.This article includes2tables,7figures and46references.