de la Fuente Ricardo, Abad José Luis, García-Castro Javier, Fernández-Miguel Gemma, Petriz Jordi, Rubio Daniel, Vicario-Abejón Carlos, Guillén Pedro, González Manuel A, Bernad Antonio
Department of Immunology and Oncology, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Exp Cell Res. 2004 Jul 15;297(2):313-28. doi: 10.1016/j.yexcr.2004.02.026.
To test the hypothesis that dedifferentiated adult human cartilage chondrocytes (HAC) are a true multipotent primitive population.
Studies to characterize dedifferentiated HAC included cell cycle and quiescence analysis, cell fusion, flow-FISH telomere length assays, and ABC transporter analysis. Dedifferentiated HAC were characterized by flow cytometry, in parallel with bone marrow mesenchymal stem cells (MSC) and processed lipoaspirate (PLA) cells. The in vitro differentiation potential of dedifferentiated HAC was studied by cell culture under several inducing conditions, in multiclonal and clonal cell populations.
Long-term HAC cultures were chromosomically stable and maintained cell cycle dynamics while showing telomere shortening. The phenotype of dedifferentiated HAC was quite similar to that of human bone marrow MSC. In addition, this population expressed human embryonic stem cell markers. Multiclonal populations of dedifferentiated HAC differentiated to chondrogenic, osteogenic, adipogenic, myogenic, and neurogenic lineages. Following VEGF induction, dedifferentiated HAC expressed characteristics of endothelial cells, including AcLDL uptake. A total of 53 clonal populations of dedifferentiated HAC were efficiently expanded; 17 were able to differentiate to chondrogenic, osteogenic, and adipogenic lineages. No correlation was observed between telomere length or quiescent population and differentiation potential in the clones assayed.
Dedifferentiated HAC should be considered a human multipotent primitive population.
验证去分化的成人人类软骨细胞（HAC）是真正的多能原始细胞群这一假说。
用于鉴定去分化HAC的研究包括细胞周期和静止分析、细胞融合、流式荧光原位杂交端粒长度测定以及ABC转运蛋白分析。通过流式细胞术对去分化HAC进行特征分析，并与骨髓间充质干细胞（MSC）和处理过的脂肪抽吸物（PLA）细胞进行平行比较。在多种诱导条件下，对多克隆和克隆细胞群体进行细胞培养，研究去分化HAC的体外分化潜能。
长期的HAC培养物染色体稳定，维持细胞周期动态，同时显示端粒缩短。去分化HAC的表型与人类骨髓MSC非常相似。此外，该细胞群表达人类胚胎干细胞标志物。去分化HAC的多克隆群体可分化为软骨、成骨、成脂、成肌和神经谱系。在VEGF诱导后，去分化HAC表现出内皮细胞特征，包括摄取乙酰化低密度脂蛋白。总共53个去分化HAC的克隆群体得到有效扩增；其中17个能够分化为软骨、成骨和成脂谱系。在所检测的克隆中，未观察到端粒长度或静止细胞群与分化潜能之间的相关性。
去分化HAC应被视为人类多能原始细胞群。
