A cell line was developed from the transplantable Swarm rat chondrosarcoma (RCS) and has been maintained in continuous monolayer tissue culture for a number of years. This long term‐cultured (LTC) cell line exhibits the morphological and biochemical characteristics of chondrocytes and resembles the RCS tumor by electron and light microscopy. The cell line differs from the original tumor cells in that about 90% of the sulfated macromolecules are retained in the LTC extracellular matrix as compared to 30% by primary cultures of cells from the RCS tumor. An interesting and useful feature of this cell line is that it contains clonal populations of cells which differ in the quality and quantity of matrix produced. Two such clones serve to illustrate the diversity of cell types within the LTC cell line. One termed Rex accumulates an intensely staining matrix around it, while the other, Ng, accumulates a matrix, that remains virtually unstained. The chondrocytic nature and ease of cloning make these cells ideal for biochemical analysis of the chondrocyte and its extracellular matrix. J. Cell. Biochem. 89: 992–1004, 2003. © 2003 Wiley‐Liss, Inc.