Summary
Growth cartilage (GC) cells of young rabbits were cultured in vitro and their homogenates were injected into mice. Hybridomas were prepared by the cell fusion technique between the myeloma cells and the spleen cells of the immunized mice. Monoclonal antibodies (MoAbs) were produced by the hybridomas in the peritoneal cavities of the mice, and some of these, temporarily named MoAbs A, B, D, N, P, and S, were studied. The localization of the antigens of each of the MoAbs in the GC or adjacent resting cartilage (RC) was examined by indirect fluorescent antibody staining. The molecular weight of the antigens was examined by immunoblot staining after SDS-polyacrylamide gel electrophoresis: MoAb A and MoAb N stained RC cells and GC cells, except calcified GC. MoAb B stained the hypertrophic and calcified GC, and matrices in the RC and proliferating GC. MoAb D stained the calcified GC. MoAb P and MoAb S stained the RC cells and the matrices in the GC, intensively in the hypertrophic GC and perichondrium. The molecular weights of the antigens of MoAbs A, P, and S were 40–70 KD, 35–40 KD and 30 KD, respectively.
Résumé
Des cellules de cartilage de croissance de jeunes lapins ont été cultivées in vitro et leurs homogénats ont été injectés à la souris. Des hybridomes ont été préparés par la technique de fusion des cellules de myélome et des cellules de la rate de souris immunisée. Des anticorps monoclonaux (MoAbs) ont été produits par les hybridomes dans la cavité péritonéale de la souris et certains d'entre eux, provisoirement appelés MoAbs A, B, D, N, P et S, ont été étudiés. La localisation des antigènes de chaque MoAbs dans le cartilage de croissance ou dans le cartilage au repos avoisinant a été recherchée par coloration fluorescente indirecte des anticorps. Le poids moléculaire des antigènes a été évalué par coloration immunogène après électrophorèse au gel polycrylamide SDS. Le MoAb A et le MoAb N ont coloré les cellules du cartilage au repos et les cellules du cartilage de croissance à l'exception du cartilage de croissance calcifié. Le MoAb B a coloré le cartilage de croissance hypertrophique et calcifié ainsi que les matrices du cartilage au repos et le cartilage de croissance proliférant. Le MoAb D a coloré le cartilage de croissance calcifié. Le MoAb P et le MoAb S ont coloré les cellules du cartilage au repos et les matrices dans le cartilage de croissance, de manière particulièrement intensive dans le cartilage de croissance hypertrophique et le périchondre. Le poids moléculaire de l'antigène des MoAbs A, P et S s'est trouvé être respectivement 40–70 KD, 35–40 KD et 30 KD.
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Supported by Japan Orthopaedics and Traumatology Foundation, Inc. (JOTF), Grant No. 0021
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Okihana, H., Shimomura, Y. Production and characterization of monoclonal antibodies against rabbit growth cartilage. International Orthopaedics 14, 321–327 (1990). https://doi.org/10.1007/BF00178767
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DOI: https://doi.org/10.1007/BF00178767