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Protective Action of Mucopolysaccharides on Dog Kidney Cell Line MDCK in Meniscus-Gradient Culture¹

Cancer Bioassay Laboratory, Department of Pathology, The Medical College of Pennsylvania, Philadelphia, Pennsylvania 19129

The effect of heparin, chondroitin sulfate, hyaluronic acid, and N-acetylneuraminic acid on the growth of Maden Darby canine kidney cells in meniscus-gradient culture was investigated.

Cell suspensions (200,000 cells/ml of 30% calf serum in Eagle's minimal essential medium) were inoculated into small Leighton tubes containing 11- x 35-mm coverslips, and the tubes were incubated in horizontal position. After 1 or 2 days, when the cells had grown to confluency, they were fed with 2 ml of medium containing either heparin, chondroitin sulfate, hyaluronic acid, or N-acetylneuraminic acid solution (0.05% of the substance added to the medium). Then all tubes were placed in a vertical position. The medium was replaced two or three times weekly.

After 5 days, marked differences were observed between the control and the treated cultures in cell viability and attachment. The cells degenerated and detached in the bottom area of the control tubes, leaving a narrow viable zone 5 mm wide just below the surface of the medium. This degeneration was, however, prevented for as long as 2 weeks when the mucopolysaccharides were added to the medium. In the treated groups the cells were viable even to a depth of 14 mm, and mitotic figures were observed at the bottom of the tube. In the heparin-treated group many blisters could be observed.

This result indicates that mucopolysaccharides may play a role in protecting and sustaining the viability and coordinated transport function of Maden Darby canine kidney cells in a microenvironment that otherwise is not compatible with viability.

¹ Supported by NIH-NCI Research Grants CA 13219, CA 14137, and Contract G-72-3858.

² On leave from the Department of Pathology, Aichi-Gakuin University, School of Dentistry, Nagoya, Japan. Reprint requests to any of the authors should be addressed to Cancer Bioassay Laboratory, Department of Pathology, The Medical College of Pennsylvania, 3300 Henry Avenue, Philadelphia, Pa. 19129.

Received 7/ 5/73. Accepted 10/11/73.