OBJECTIVE: To investigate the contribution of clinical ex vivo expansion protocols to replicative aging of human chondrocytes. METHODS: Primary human chondrocytes were cultured as monolayers after isolation from 7 articular cartilage specimens. Cells were passaged corresponding to 12-19 cell population doublings (cpd). Aliquots of the cells were collected from each passage and analyzed for telomere length and telomerase activity. RESULTS: The rate of telomere shortening was heterogeneous, ranging from 147 to 431 bp/cpd (mean +/- SD 305 +/- 122). Telomerase activity was detected at various time points during passaging in 5 of 7 primary chondrocytes analyzed, but not in native human articular cartilage specimens. According to our data, an 8-10-fold ( approximately 3 cpd) ex vivo expansion of articular chondrocytes, as typically performed for transplantation procedures, leads to telomere erosion in the range of 900 bp. This is comparable with 30 years of aging based on the in vivo rate of telomere shortening of 30 bp/year recently found in chondrocytes. CONCLUSION: If telomere shortening is an important determinant of aging in human articular cartilage, an additional telomere loss due to ex vivo expansion might affect the incidence or time of onset of age-related cartilage disorders. However, given the limited extent of expansion performed in the clinical setting to date, a significant telomere-mediated increase in the risk of malignant transformation or replicative exhaustion of the transplanted cells seems unlikely.
OBJECTIVE: To investigate the contribution of clinical ex vivo expansion protocols to replicative aging of human chondrocytes. METHODS: Primary human chondrocytes were cultured as monolayers after isolation from 7 articular cartilage specimens. Cells were passaged corresponding to 12-19 cell population doublings (cpd). Aliquots of the cells were collected from each passage and analyzed for telomere length and telomerase activity. RESULTS: The rate of telomere shortening was heterogeneous, ranging from 147 to 431 bp/cpd (mean +/- SD 305 +/- 122). Telomerase activity was detected at various time points during passaging in 5 of 7 primary chondrocytes analyzed, but not in native humanarticular cartilage specimens. According to our data, an 8-10-fold ( approximately 3 cpd) ex vivo expansion of articular chondrocytes, as typically performed for transplantation procedures, leads to telomere erosion in the range of 900 bp. This is comparable with 30 years of aging based on the in vivo rate of telomere shortening of 30 bp/year recently found in chondrocytes. CONCLUSION: If telomere shortening is an important determinant of aging in humanarticular cartilage, an additional telomere loss due to ex vivo expansion might affect the incidence or time of onset of age-related cartilage disorders. However, given the limited extent of expansion performed in the clinical setting to date, a significant telomere-mediated increase in the risk of malignant transformation or replicative exhaustion of the transplanted cells seems unlikely.
Authors: Karsten Schrobback; Jana Wrobel; Dietmar W Hutmacher; Tim B F Woodfield; Travis J Klein Journal: Tissue Eng Part A Date: 2013-02-19 Impact factor: 3.845
Authors: Minjie Zhang; Sriniwasan B Mani; Yao He; Amber M Hall; Lin Xu; Yefu Li; David Zurakowski; Gregory D Jay; Matthew L Warman Journal: J Clin Invest Date: 2016-07-18 Impact factor: 14.808