David H Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Pharm Res. 2011 Jun;28(6):1395-405. doi: 10.1007/s11095-011-0445-2. Epub 2011 Apr 15.
Chondrocytes lose their ability to produce cartilaginous matrix during multiplication in culture through repeated passages, resulting in inferior tissue phenotype. To overcome the limited amount of primary chondrocytes, we aimed to determine the optimal culture condition for in vitro/in vivo cartilage regeneration using human adipose-derived mesenchymal stem cells (AMSCs).
To evaluate the effects exerted by the chondrocytic culture condition on AMSC, we utilized chondrocyte conditioned medium (CM) and/or co-culture methods to prime and differentiate AMSCs. We evaluated ultimate in vivo engineered cartilage with primed AMSCs with that of chondrocytes. To examine the link between conditioned factors and proliferation/differentiation, cell cycle progression of AMSCs were examined using 5-ethynyl-2'-deoxyuridine (EdU), and gene expression was monitored.
We report that AMSCs can be stimulated to become chondrogenic cells when expanded with chondrocyte CM. Polymeric scaffolds co-seeded with CM- expanded AMSCs and primary chondrocytes resulted in in vivo cartilaginous tissues with similar biochemical content to constructs seeded with chondrocytes alone.
These results indicate that chondrocyte CM consists of suitable morphogenetic factors that induce the chondrogenic priming of AMSCs for cartilage tissue engineering.
在培养过程中,软骨细胞通过反复传代而失去增殖过程中产生软骨基质的能力,导致组织表型下降。为了克服原代软骨细胞数量有限的问题,我们旨在确定使用人脂肪间充质干细胞(AMSCs)进行体外/体内软骨再生的最佳培养条件。
为了评估软骨细胞培养条件对 AMSC 的影响,我们采用软骨细胞条件培养基(CM)和/或共培养方法对 AMSCs 进行诱导和分化。我们将经过诱导的 AMSCs 与软骨细胞的最终体内工程化软骨进行比较。为了研究条件因子与增殖/分化之间的关系,我们使用 5-乙炔基-2'-脱氧尿苷(EdU)检测 AMSCs 的细胞周期进程,并监测基因表达。
我们报告称,当用软骨细胞 CM 扩增时,AMSCs 可以被刺激成为软骨细胞。与单独用软骨细胞接种的构建体相比,与 CM 扩增的 AMSCs 和原代软骨细胞共接种的聚合物支架在体内产生了具有相似生化含量的软骨组织。
这些结果表明,软骨细胞 CM 中含有合适的形态发生因子,可诱导 AMSCs 的软骨起始,用于软骨组织工程。