Department of Trauma and Reconstructive Surgery, University of Regensburg, Franz-Josef-Strass-Allee 11, Regensburg 93042, Germany.
J Orthop Res. 2010 Mar;28(3):354-60. doi: 10.1002/jor.21007.
Decreasing replicative potential and dedifferentiation of articular chondrocytes during expansion in cell culture are essential limitations for tissue engineering and cell therapy approaches. Telomeres and telomerase play a key role in cell development, aging, and tumorigenesis. There is evidence that growth factors are involved in regulating telomerase activity. Therefore, the objective was to evaluate the effect of selected growth factors on telomere biology of serially passaged chondrocytes. Human articular chondrocytes were isolated from cartilage of three patients undergoing total knee arthroplasty. The chondrocytes were cultured in monolayer with the growth factors PDGF-BB, TGF-beta1, and bFGF. Telomere length was measured by telomere restriction fragment length assay, and telomerase activity was determined by quantifying the gene expression of its catalytic subunit hTERT by rtPCR. Chondrocytes cultured with PDGF-BB and TGF-beta1 showed a significantly higher proliferation rate than control cells. None of the growth factor cultures revealed an accelerated rate of telomere shortening. Telomerase was not expressed in significant amounts in any of the chondrocyte cultures. Growth factor treatment of chondrocyte cell cultures for cell therapy purposes can be regarded as safe in terms of telomere biology.
在细胞培养中扩增时，关节软骨细胞的复制潜能和去分化降低是组织工程和细胞治疗方法的重要限制。端粒和端粒酶在细胞发育、衰老和肿瘤发生中起着关键作用。有证据表明，生长因子参与调节端粒酶活性。因此，本研究的目的是评估选定的生长因子对传代软骨细胞端粒生物学的影响。从 3 名接受全膝关节置换术的患者的软骨中分离出关节软骨细胞。软骨细胞在含有 PDGF-BB、TGF-β1 和 bFGF 的生长因子的单层中培养。通过端粒限制片段长度分析测量端粒长度，通过定量分析其催化亚基 hTERT 的基因表达来确定端粒酶活性。与对照组细胞相比，用 PDGF-BB 和 TGF-β1 培养的软骨细胞增殖速度明显更快。在任何软骨细胞培养物中都没有发现端粒缩短的加速。在任何软骨细胞培养物中都没有检测到端粒酶的大量表达。就端粒生物学而言，生长因子处理软骨细胞培养物用于细胞治疗是安全的。
