Investigation Of The Effect Of Bone Marrow Mesenchymal Stem Cells On The Chronic Idiopathic Thrombocytopenic Purpura In Vitro

Objective: To study the feasibility of allogeneic bone marrow mesenchymal stem cells for the treatment of chronic idiopathic thrombocytopenic purpura in vitro .Methods: 1. BM-MSCs were isolated from normal human and cultured for proliferating to passage cells; cell morphology and growth characteristics of BM-MSCs were observed under inverted microscope.2. PBMNCs were isolated from chronic idiopathic thrombocytopenic purpura patients, and were purificated and cultured 3. Allogeneic BM-MSCs and PBMNCs stimulated by pokeweed of cITP patients were coculture; lymphocyte proliferation was tested with MTT assay before and after cocultivation; change of CD19 + CD5 + B1 lymphocytes percentage in lymphocytes of each groups test by flow cytometry; the PAIgG content of each group of cell culture supernatant was test by ELISA. 4. 50%, 25% or no BM-MSCs–LCM was added in normal bone marrow megakaryocyte colony culture system; the influence of BM-MSCs-LCM on the normal bone marrow megakaryocyte colony formation was observed by counting the number of CFU-MK of each groups.Results:1.Normal human bone marrow MSCs adhered the bottom of the flask and were fusiform shape, could continuously passage in vitro;2. Compared with control group ,the proliferation of PBMNCs co-cultured with BM-MSCs was significantly inhibited ( P<0.01 ) ; 3.Compared with control group, Allogeneic BM-MSCs could significantly reduce the percentage of CD +19 CD5 + B1 lymphocytes in total lymphocytes of peripheral blood of cITP patient( P<0.01) , could significantly inhibited the secretion of PAIgG by lymphocytes of cITP patient( P<0.01) (PBMNCs: BM-MSCs=10:1);4.We Found that BM-MSCs-LCM have no significant effect on the formation of CFU-MK in vitro( P>0.05)。Conclusion:1.Normal human bone marrow mesenchymal stem cells could significantly inhibit the proliferation stimulated by pokeweed mitogen of peripheral blood lymphocytes in patients with cITP; could significantly inhibit the the secretion of PAIgG by B lymphocytes in patients with cITP; could significantly reduce the percentage of CD19+ CD5 + B1 lymphocytes in total lymphocytes of peripheral blood of cITP patient.2.BM-MSCs-LCM have no significant effect on the formation of normal human bone marrow CFU-MK in vitro. 3. Our extraneous experiments showed that BM-MSCs have the possibility in the treatment of cITP patient.