人骨髓间充质干细胞中Toll样受体的表达特征

本研究探讨健康供体骨髓源性间充质干细胞（BM—MSC）中Toll样受体（TLR）的表达特征，，用Ficoll法分离培养健康供体的BM—MSC；流式细胞术（FCM）检测BM—MSC中CD34、CD45、HLA—DR、CD44和CD71的表达；免疫细胞化学法检测爬片BM—MSC细胞中CD71的表达；应用茜素红染色和油红染色检测BM—MSC细胞成骨及成脂诱导情况；半定量RT—PCR法检测TLR1—10mRNA的表达。结果表明，体外分离培养的BM—MSC中CD34、CD45和HLA—DR表达呈阴性．CD44和CD71表达呈阳性；爬片BM-MSC细胞中CD71的表达呈阳性。BM—MSC经成骨和成脂诱导后出现钙沉积和脂滴，茜素红染色和油红染色均呈阳性。BM—MSC中TLR1—10mRNA均有表达，其中TLR2、TLR3、TLR4、TI，R7、TLR8、TLR9均高表达，TLR1、TLR5、TLR6、TLR10均低表达。结论：从健康供体骨髓中分离培养的BM—MSC表达不同水平的TLR1-10111RNA。

Expression of Toll-like Receptors in Human Bone Marrow Mesenchemal Stem Cells

The aim of this study was to explore the characteristics of Toll-like receptor expression in mesenchemal stern cells derived from bone marrow of healthy donor （BM-MSCs）. BM-MSCs were isolated from bone marrow of healthy donor by Ficoll method. Expressions of CD34, CIM5, HLA-DR, CD44 and CD71 in BM-MSCs were detected by flow cytometry. CD71 in BM-MSCs was assayed by immunocytochemistry. The adipocyte and osteoblast induction of BM-MSCs were detected by alizarin red stain and oil red stain respectively. TLRI-10 mRNA levels in BM-MSCs were evaluated by semiquantitative RT-PCR. The results showed that expressions of CD34, CD45 and HLA-DR in BM-MSC were negative while the expressions of CD44 and CD71 were positive. CD71 in BM-MSCs was possitive. After induced by osteoblast and adiopocyte inductor, BM-MSCs were positive for alizarin red staining and oil red staining respectively. All of TLRI-10 mRNA were found in BM-MSCs with high expression levels of TLR2, TLR3, TLR4, TLR7, TLR8, TLR9 and low expression levels of TLR1, TLR5, TLR6, TLR10. In conclusion, different levels of TLR 1-10 mRNA were expressed in BM-MSCs of healthy donor.