Abstract
Objective To investigate the effect of recombinant human erythropoietin (rhEPO) on proliferation of human bone marrow-derived mesenchymal stem cells (MSCs) in culture and its probable molecular mechanism. Methods The aspirates of the bone marrow from healthy volunteers were seeded in culture medium. Then MSCs were isolated by its character adhering to the plastic. After three passages in culture, MSCs were identified by morphological features, cell surface antigens and differentiation into multi-lineages. Then P3-MSCs were treated with different concentrations of rhEPO (0.5, 1, 5, 10, or 50 U/ml). And proliferation of MSCs was measured by MTT assay. Subsequently, EPO receptor (EPOR) was blocked and MTT was performed for another time. What's more, immuofluresence cytochemical staining was used to detect the expression of EPOR. And the proteins of signal pathway were examined by Western blot. Results The P3 bone morrow-derived adherent cells were positive for CD90 and CD105, while negative for CD34 and CD45, and could differentiate into adipocytes, osteocytes and chondrocytes in induction media. MTT assays showed that the absorbance (A value) in EPO-treated groups was significantly higher than in control group, and 10 U/ml EPO-treated group achieved the most predominant effects. However, the A value was reduced dramatically when the specific anti-EPOR antibody (anti-EPOR Ab) was added with rhEPO. Compared with the control group, all those differences were statistically significant (P<0.01). In addition, P3-MSCs were positive for EPOR. And the expression of EPOR, phospho-Janus kinase2 (pJAK2) and phospho-signal transducer and activator of transcription (pSTAT-5) was significantly higher in 10 U/ml EPO-treated group than in the control group. Conclusion Erythropoietin can promote proliferation of human bone marrow MSCs in vitro, which is mediated by EOPOR, and related with JAK2-STAT-5 signal pathway. Key words: Erythropoietin, recombinant; Bone marrow; Mesenchymal stem cells; Cell proliferation
Published Version
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