Larson Christopher M, Kelley Scott S, Blackwood A Denene, Banes Albert J, Lee Greta M
Department of Orthopedics, University of North Carolina School of Medicine, Chapel Hill, NC, USA.
Matrix Biol. 2002 Jun;21(4):349-59. doi: 10.1016/s0945-053x(02)00026-4.
The interaction of the cell with its surrounding extracellular matrix (ECM) has a major effect on cell metabolism. We have previously shown that chondrons, chondrocytes with their in vivo-formed pericellular matrix, can be enzymatically isolated from articular cartilage. To study the effect of the native chondrocyte pericellular matrix on ECM production and assembly, chondrons were compared with chondrocytes isolated without any pericellular matrix. Immediately after isolation from human cartilage, chondrons and chondrocytes were centrifuged into pellets and cultured. Chondron pellets had a greater increase in weight over 8 weeks, were more hyaline appearing, and had more type II collagen deposition and assembly than chondrocyte pellets. Minimal type I procollagen immunofluorescence was detected for both chondron and chondrocyte pellets. Chondron pellets had a 10-fold increase in proteoglycan content compared with a six-fold increase for chondrocyte pellets over 8 weeks (P<0.0001). There was no significant cell division for either chondron or chondrocyte pellets. The majority of cells within both chondron and chondrocyte pellets maintained their polygonal or rounded shape except for a thin, superficial edging of flattened cells. This edging was similar to a perichondrium with abundant type I collagen and fibronectin, and decreased type II collagen and proteoglycan content compared with the remainder of the pellet. This study demonstrates that the native pericellular matrix promotes matrix production and assembly in vitro. Further, the continued matrix production and assembly throughout the 8-week culture period make chondron pellet cultures valuable as a hyaline-like cartilage model in vitro.
细胞与其周围细胞外基质(ECM)的相互作用对细胞代谢有重大影响。我们之前已经表明,软骨粒,即带有其体内形成的细胞周围基质的软骨细胞,可以通过酶法从关节软骨中分离出来。为了研究天然软骨细胞周围基质对ECM产生和组装的影响,将软骨粒与未带有任何细胞周围基质而分离出的软骨细胞进行了比较。从人软骨中分离后,立即将软骨粒和软骨细胞离心成沉淀并进行培养。与软骨细胞沉淀相比,软骨粒沉淀在8周内重量增加更多,外观更呈透明状,并且有更多的II型胶原蛋白沉积和组装。对于软骨粒和软骨细胞沉淀,均检测到极少的I型前胶原免疫荧光。与软骨细胞沉淀在8周内增加6倍相比,软骨粒沉淀的蛋白聚糖含量增加了10倍(P<0.0001)。软骨粒和软骨细胞沉淀均无明显的细胞分裂。软骨粒和软骨细胞沉淀内的大多数细胞保持其多边形或圆形形状,除了一层薄薄的、表面呈扁平状的边缘细胞。该边缘类似于富含I型胶原蛋白和纤连蛋白的软骨膜,与沉淀的其余部分相比,II型胶原蛋白和蛋白聚糖含量降低。本研究表明,天然细胞周围基质在体外促进基质的产生和组装。此外,在整个8周培养期内持续的基质产生和组装使软骨粒沉淀培养作为一种体外透明样软骨模型具有重要价值。