Abstract
We developed a novel cloning method of synovial lining cells from the bovine metacarpophalangeal joint. The synovial lining cells were mechanically dissected without contamination of cells derived from the sublining layer. Nine clones were established out of 46 trials, a success-rate of about 20%. The cell type of the established clone was determined with transmission and scanning electron microscopy. With the criteria for B-and A-cells, 5 clones could be classified as B-cells and 3 clones as A-cells with one clone unidentified. Using a patch-clamp, we further studied the excitability and distribution of ion channels in 1- to 5-month clone-cultured cells, and found that 1) B-cells lost electrical excitability, 2) the resting membrane potential was kept normal in both B- and A-cells, and 3) B- and A-cells showed a tendency to resemble each other in their distribution of channels, which originally had been different. These results show that while cells normally increase in number and their morphological characteristics are retained, their electrophysiological nature is apparently altered in long-term clone-cultured cells.
