Gene-induced chondrogenesis of primary mesenchymal stem cells in vitro
Gene-induced chondrogenesis of primary mesenchymal stem cells in vitro
复制标题
DOI：
10.1016/j.ymthe.2005.03.024
复制
发表时间：
2005-08-01
影响因子：
12.4
通讯作者：
Ghivizzani, SC
中科院分区：
文献类型：
--
作者：
Palmer, GD;Steinert, A;Ghivizzani, SC
成体间充质干细胞（MSCs）在某些生长因子的刺激下具有分化为各种结缔组织如软骨和骨的能力。然而，当这些蛋白质通过基因转移传递时，对这些细胞进行软骨形成的能力知之甚少。在这项研究中，我们研究了原代骨髓来源的间充质干细胞在聚合培养物中的软骨形成，这些细胞在用编码软骨形成生长因子的腺病毒载体进行遗传修饰后。我们发现，腺病毒介导的TGF-β 1和BMP-2的表达，而不是IGF-1，诱导骨髓间充质干细胞的软骨形成，甲苯胺蓝异染和II型胶原的免疫组化检测证明。软骨形成与表达蛋白的水平和持续时间相关，并且在表达10-100 ng/ml转基因产物的聚集体中最强。所有聚集体中的转基因表达是高度瞬时的，在7天后显示出显著降低。在修饰为表达>100 ng/ml TGF-β 1或BMP-2的聚集体中，软骨形成受到抑制;然而，发现这部分是由于暴露于高腺病毒载量的抑制作用。我们的研究结果表明，这些参数是适应基因转移技术诱导间充质干细胞软骨形成的重要功能考虑因素。
Adult mesenchymal stem cells (MSCs) have the capacity to differentiate into various connective tissues such as cartilage and bone following stimulation with certain growth factors. However, less is known about the capacity of these cells to undergo chondrogenesis when these proteins are delivered via gene transfer. In this study, we investigated chondrogenesis of primary, bone marrow-derived MSCs in aggregate cultures following genetic modification with adenoviral vectors encoding chondrogenic growth factors. We found that adenoviral-mediated expression of TGF-beta 1 and BMP-2, but not IGF-1, induced chondrogenesis of MSCs as evidenced by toluidine blue metachromasia and immunohistochemical detection of type II collagen. Chondrogenesis correlated with the level and duration of expressed protein and was strongest in aggregates expressing 10-100 ng/ml transgene product. Transgene expression in all aggregates was highly transient, showing a marked decrease after 7 days. Chondrogenesis was inhibited in aggregates modified to express >100 ng/ml TGF-beta 1 or BMP-2; however, this was found to be partly due to the inhibitory effect of exposure to high adenoviral loads. Our findings indicate that parameters such as these are important functional considerations for adapting gene transfer technologies to induce chondrogenesis of MSCs.