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The Late Osteoblast/Preosteocyte Cell Line MLO-A5 Displays Mesenchymal Lineage Plasticity In Vitro and In Vivo

机译:晚期成骨细胞/前成骨细胞系MLO-A5在体外和体内显示间充质谱系可塑性

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摘要

The process of osteoblast switching to alternative mesenchymal phenotypes is incompletely understood. In this study, we tested the ability of the osteoblast/preosteocyte osteogenic cell line, MLO-A5, to also differentiate into either adipocytes or chondrocytes. MLO-A5 cells expressed a subset of skeletal stem cell markers, including Sca-1, CD44, CD73, CD146, and CD166. Confluent cultures of cells underwent differentiation within 3 days upon the addition of osteogenic medium. The same cultures were capable of undergoing adipogenic and chondrogenic differentiation under lineage-appropriate culture conditions, evidenced by lineage-specific gene expression analysis by real-time reverse-transcription-PCR, and by Oil Red O and alcian blue (pH 2.5) staining, respectively. Subcutaneous implantation of MLO-A5 cells in a gel foam into NOD SCID mice resulted in a woven bone-like structure containing embedded osteocytes and regions of cartilage-like tissue, which stained positive with both alcian blue (pH 2.5) and safranin O. Together, our findings show that MLO-A5 cells, despite being a strongly osteogenic cell line, exhibit characteristics of skeletal stem cells and display mesenchymal lineage plasticity in vitro and in vivo. These unique characteristics suggest that this cell line is a useful model with which to study aging and disease-related changes to the mesenchymal lineage composition of bone.
机译:成骨细胞转换为间充质表型的过程尚不完全清楚。在这项研究中,我们测试了成骨细胞/前骨细胞成骨细胞系MLO-A5分化为脂肪细胞或软骨细胞的能力。 MLO-A5细胞表达了骨骼干细胞标记的子集,包括Sca-1,CD44,CD73,CD146和CD166。加入成骨培养基后三天内,细胞融合培养物开始分化。相同的培养物能够在适合谱系的培养条件下进行成脂和软骨分化,这通过实时逆转录PCR的谱系特异性基因表达分析以及油红O和阿尔辛蓝(pH 2.5)染色来证明,分别。将MLO-A5细胞以凝胶泡沫的形式皮下植入NOD SCID小鼠体内,形成了一个编织的骨样结构,其中包含包埋的骨细胞和软骨样组织区域,并被阿尔辛蓝(pH 2.5)和番红素O染色呈阳性。 ,我们的发现表明,尽管MLO-A5细胞是强成骨细胞系,但在体外和体内均表现出骨骼干细胞的特征并显示间充质谱系可塑性。这些独特的特征表明,这种细胞系是研究骨骼间质谱系组成的衰老和疾病相关变化的有用模型。

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