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植入前培养软骨细胞中端粒长度的研究

Study of Telomere Length in Preimplanted Cultured Chondrocytes.

作者信息

López-Alcorocho Juan Manuel, Guillén-Vicente Isabel, Rodríguez-Iñigo Elena, Guillén-Vicente Marta, Fernández-Jaén Tomás Fernando, Caballero Rosa, Casqueiro Mercedes, Najarro Pilar, Abelow Steve, Guillén-García Pedro

机构信息

1 Research Unit, Clínica Cemtro, Madrid, Spain.

2 Life Length, Madrid, Spain.

出版信息

Cartilage. 2019 Jan;10(1):36-42. doi: 10.1177/1947603517749918. Epub 2018 Jan 11.

Abstract

DESIGN

In the process of cell division, the extremes of the eukaryotic chromosomes are progressively shortening, and this phenomenon is related to cell degeneration and senescence. The treatment of cartilage lesions with autologous chondrocytes implies that cells proliferate in an artificial environment. We have studied the viability of cultured chondrocytes after measurement of their telomere length before implantation.

METHODS

Articular cartilage biopsies (B1, B2, and B3) were obtained from 3 patients (2 males and 1 female) with knee cartilage defects, who were going to be treated with chondrocyte implantation. Chondrocytes were cultured in DMEM with autologous serum. After the third passage, an aliquot of 1 million cells was removed to estimate the telomere length and the remaining cells were implanted. Telomere length was measured by quantitative fluorescent in situ hybridization (Q-FISH). Patients' clinical outcome was determined preoperatively, and 12 and 24 months postimplantation with the International Knee Documentation Committee (IKDC) questionnaire.

RESULTS

After chondrocyte implantation, IKDC score doubled at 12 and 24 months with regard to the basal value. After 3 passages, chondrocytes were cultured for a mean of 45.67 days, the mean duplication time being 4.53 days and the mean number of cell divisions being 10.04 during the culture period. The 20th percentile of telomere lengths were 6.84, 6.96, and 7.06 kbp and the median telomere lengths 10.30, 10.47, and 10.73 kbp, respectively. No significant correlation was found between IKDC score and telomere length.

CONCLUSION

Culturing autologous chondrocytes for implantation is not related to cell senescence in terms of telomere length.

摘要

设计

在细胞分裂过程中,真核染色体的末端会逐渐缩短,这种现象与细胞退化和衰老有关。用自体软骨细胞治疗软骨损伤意味着细胞在人工环境中增殖。我们在植入前测量了培养的软骨细胞的端粒长度,以研究其活力。

方法

从3例(2男1女)膝关节软骨缺损患者身上获取关节软骨活检样本(B1、B2和B3),这些患者将接受软骨细胞植入治疗。软骨细胞在含自体血清的DMEM中培养。传代三次后,取出100万个细胞的等分试样以估计端粒长度,其余细胞进行植入。通过定量荧光原位杂交(Q-FISH)测量端粒长度。术前以及植入后12个月和24个月,使用国际膝关节文献委员会(IKDC)问卷确定患者的临床结果。

结果

软骨细胞植入后,IKDC评分在12个月和24个月时相对于基础值翻倍。传代三次后,软骨细胞平均培养45.67天,平均倍增时间为4.53天,培养期间平均细胞分裂次数为10.04次。端粒长度的第20百分位数分别为6.84、6.96和7.06 kbp,端粒长度中位数分别为10.30、10.47和10.73 kbp。未发现IKDC评分与端粒长度之间存在显著相关性。

结论

就端粒长度而言,培养用于植入的自体软骨细胞与细胞衰老无关。

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