Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, Neston, Cheshire, United Kingdom.
Tissue Eng Part A. 2011 Jul;17(13-14):1801-7. doi: 10.1089/ten.TEA.2010.0579. Epub 2011 Apr 18.
Marker genes are used to monitor chondrogenic differentiation, but little is known about the turnover of their mRNA during this process. We set out to measure the half life of mRNA encoding the transcription factor SOX9, an important marker of chondrocytic phenotype. We dedifferentiated human articular chondrocytes in monolayer culture before placing them in chondrogenic three-dimensional pellet cultures. At the same time, we induced chondrocytic differentiation of human bone marrow-derived mesenchymal stem cells under the same three-dimensional conditions. Pellets were cultured in standard chondrogenic media with and without BMP7. We found that SOX9 mRNA half life exhibited an inverse correlation with total SOX9 mRNA levels in both dedifferentiating human articular chondrocytes and chondrogenic pellet cultures. There was no evidence for a specific effect of BMP7 on SOX9 mRNA decay. Our findings provide an insight into a level of gene control rarely explored in regenerative medicine, which could be important in the optimization of in vitro cartilage production.
标记基因被用于监测软骨分化，但在这个过程中，关于它们的 mRNA 周转率的信息知之甚少。我们旨在测量编码转录因子 SOX9 的 mRNA 的半衰期，SOX9 是软骨细胞表型的重要标志物。我们在单层培养中将人关节软骨细胞去分化，然后将其置于软骨形成的三维微球培养中。同时，我们在相同的三维条件下诱导人骨髓间充质干细胞的软骨细胞分化。微球在有和没有 BMP7 的标准软骨形成培养基中培养。我们发现，在去分化的人关节软骨细胞和软骨形成微球培养物中，SOX9 mRNA 半衰期与总 SOX9 mRNA 水平呈反比关系。没有证据表明 BMP7 对 SOX9 mRNA 降解有特定影响。我们的发现为再生医学中很少探索的基因控制水平提供了深入了解，这对于优化体外软骨生成可能很重要。
