在妊娠合并免疫性血小板减少症动物模型中骨髓间充质干细胞的改变#br#

Changes of bone marrow mesenchymal stem cells in animal model of immune thrombocytopenia during pregnancy

Objective?To establish a method for isolation and culture of bone marrow mesenchymal stem cells (BM-MSCs) in mice with immune thrombocytopenia (ITP) during pregnancy. Methods?Mesenchymal stem cells were isolated and cultured from mouse bone marrow of normal pregnancy, ITP and ITP pregnancy group. Cell growth and proliferation were observed under optical microscope. The immunophenotype of mesenchymal stem cells was analyzed by flow cytometry. Oil red 0 and alizarin red staining were used to evaluate the ability of lipid osteogenesis differentiation. CCk-8 test was used to detect cell proliferation. The PI test measures the cell cycle.Annexin-v /PI staining was used to detect apoptosis. Results?①BM-MSCs derived from pregnant mice with ITP group grew slowly and were irregular in shape. BM-MSCs from different sub-groups were highly expressed in stem-cell related antigens CD44, CD29 and scal-1, partially expressing CD90, but not expressing leukocyte surface antigen CD45 and lymphocyte surface antigen CDllb. They can be induced to differentiate into osteoblasts and lipoblasts in vitro. ②Compared with the normal pregnancy group, the proliferation of BM-MSCs in the ITP group and the ITP pregnancy group was significantly inhibited (P<0.05), the cell proportion in G0/G1 phase was significantly increased (P<0.05), and the apoptosis rate was significantly increased (P<0.05). Compared with the ITP group, the growth rate of BM-MSCs in the ITP pregnancy group was inhibited and the apoptosis rate was increased (P<0.05). Conclusions?The BM-MSCs derived from mice in the ITP pregnancy group showed abnormal morphology, impaired proliferation ability and excessive apoptosis.