Adkisson H Davis, Martin James A, Amendola Richard L, Milliman Curt, Mauch Kelsey A, Katwal Arbindra B, Seyedin Mitchell, Amendola Annuziato, Streeter Philip R, Buckwalter Joseph A
ISTO Technologies, Inc, St Louis, Missouri, St Louis, MO 63132, USA.
Am J Sports Med. 2010 Jul;38(7):1324-33. doi: 10.1177/0363546510361950. Epub 2010 Apr 27.
Donor-site morbidity, limited numbers of cells, loss of phenotype during ex vivo expansion, and age-related decline in chondrogenic activity present critical obstacles to the use of autologous chondrocyte implantation for cartilage repair. Chondrocytes from juvenile cadaveric donors may represent an alternative to autologous cells. Hypothesis/
The authors hypothesized that juvenile chondrocyte would show stronger and more stable chondrogenic activity than adult cells in vitro and that juvenile cells pose little risk of immunologic incompatibility in adult hosts.
Controlled laboratory study.
Cartilage samples were from juvenile (<13 years old) and adult (>13 years old) donors. The chondrogenic activity of freshly isolated human articular chondrocytes and of expanded cells after monolayer culture was measured by proteoglycan assay, gene expression analysis, and histology. Lymphocyte proliferation assays were used to assess immunogenic activity.
Proteoglycan content in neocartilage produced by juvenile chondrocytes was 100-fold higher than in neocartilage produced by adult cells. Collagen type II and type IX mRNA in fresh juvenile chondrocytes were 100- and 700-fold higher, respectively, than in adult chondrocytes. The distributions of collagens II and IX were similar in native juvenile cartilage and in neocartilage made by juvenile cells. Juvenile cells grew significantly faster in monolayer cultures than adult cells (P = .002) and proteoglycan levels produced in agarose culture was significantly higher in juvenile cells than in adult cells after multiple passages (P < .001). Juvenile chondrocytes did not stimulate lymphocyte proliferation.
These results document a dramatic age-related decline in human chondrocyte chondrogenic potential and show that allogeneic juvenile chondrocytes do not stimulate an immunologic response in vivo.
Juvenile human chondrocytes have greater potential to restore articular cartilage than adult cells, and may be transplanted without the fear of rejection, suggesting a new allogeneic approach to restoring articular cartilage in older individuals.
供区并发症、细胞数量有限、体外扩增过程中表型丧失以及软骨生成活性的年龄相关性下降，是自体软骨细胞移植用于软骨修复面临的关键障碍。来自青少年尸体供体的软骨细胞可能是自体细胞的一种替代选择。假设/目的：作者假设青少年软骨细胞在体外比成年细胞表现出更强且更稳定的软骨生成活性，并且青少年细胞在成年宿主中引发免疫不相容的风险很小。研究设计：对照实验室研究。方法：软骨样本取自青少年（<13岁）和成年（>13岁）供体。通过蛋白聚糖测定、基因表达分析和组织学检测新鲜分离的人关节软骨细胞以及单层培养后扩增细胞的软骨生成活性。采用淋巴细胞增殖试验评估免疫原性活性。结果：青少年软骨细胞产生的新软骨中的蛋白聚糖含量比成年细胞产生的新软骨高100倍。新鲜青少年软骨细胞中Ⅱ型和Ⅸ型胶原蛋白mRNA分别比成年软骨细胞高100倍和700倍。Ⅱ型和Ⅸ型胶原蛋白在天然青少年软骨和青少年细胞生成的新软骨中的分布相似。青少年细胞在单层培养中比成年细胞生长明显更快（P = 0.002），多次传代后，青少年细胞在琼脂糖培养中产生的蛋白聚糖水平显著高于成年细胞（P < 0.001）。青少年软骨细胞不会刺激淋巴细胞增殖。结论：这些结果证明人类软骨细胞的软骨生成潜能存在显著的年龄相关性下降，并表明异体青少年软骨细胞在体内不会刺激免疫反应。临床意义：青少年人类软骨细胞比成年细胞具有更大的恢复关节软骨的潜力，并且可以在无需担心排斥反应的情况下进行移植，这为老年个体恢复关节软骨提供了一种新的异体方法。
