carbocyanines and Cell-Transformation--Viral

The recent establishment of an immortalized clonal cell line of rat parotid acinar cells (2RSG) by transfecting isoproterenol-stimulated parotid cells with a plasmid vector, pSV3neo which carries the large T-antigen gene from SV40 virus, afforded the opportunity to develop a model for parotid acinar cell transplantation. Single cell suspensions of 2RSG cells labeled with a fluorescent tracer, DiI, were injected into the parotid gland or oral submucosa of allogeneic adult rats. The grafted cells survived and were functionally viable for at least 30 days. Histological sections revealed no evidence of infiltration of leukocytes or lymphocytes. Grafted cells did not form tumors. Results suggest that allogeneic parotid acinar cell transplantation is a feasible technique in the animal model.

Topics: Animals; Antigens, Polyomavirus Transforming; Carbocyanines; Cell Line, Transformed; Cell Transformation, Viral; Cell Transplantation; Clone Cells; Feasibility Studies; Fluorescent Dyes; Graft Survival; Male; Mouth Mucosa; Parotid Gland; Rats; Rats, Sprague-Dawley; Simian virus 40; Transplantation, Heterotopic; Xerostomia

We measured the lateral diffusion of the fluorescent lipid analogue dioctadecylindocarbocyanine iodide (DiI) and of membrane glycoproteins labeled with tetramethylrhodamine (TRITC) succinyl concanavalin A (SConA) via fluorescence photobleaching recovery (FPR) at selected times during a temperature downshift experiment on transformation-defective temperature-sensitive (td-ts) Rous sarcoma virus (RSV) NY68-transformed chicken embryo fibroblasts (CEF) and on identically treated CEF and RSV-transformed CEF. There were no significant differences in the lateral diffusion in DiI at any of the times measured. The lateral diffusion of TRITC-SConA on the RSV-transformed CEF, (1.32 +/- 0.12).10(-10) cm2 s-1, was approximately two times faster than that observed in normal CEF, (0.61 +/- 0.06).10(-10) cm2 s-1. In the cells undergoing RSV NY68-mediated transformation, TRITC-SConA diffusion increased over a 24-h period from a value comparable to that observed in normal CEF, (0.72 +/- 0.13).10(-10) cm2 s-1 to a value comparable to the RSV-CEF transformed cells, (1.74 +/- 0.20).10(-10) cm2 s-1. All diffusion measurements reported were made at the permissive temperature for RSV-NY68 (35 degrees C) unless stated otherwise. The changes in the lateral diffusion of TRITC-SConA occurred between the fifth and twelfth hour of the downshift course and could be associated with cytoskeletal disruption and/or fibronectin degradation, both known to occur at this time in RSV-transformed cells. To assess the contribution of extracellular matrix (ECM) degradation, SConA mobility was measured in normal and RSV-transformed cells treated with trypsin. This treatment increased SConA mobility approximately 4-fold in the normal cells relative to untreated controls and only 2-fold in the RSV-CEF transformed cells. No significant difference in SConA mobility between trypsinized spherical normal and transformed cells was apparent.

Topics: Animals; Avian Sarcoma Viruses; Carbocyanines; Cell Transformation, Viral; Cells, Cultured; Chick Embryo; Concanavalin A; Diffusion; Fibroblasts; Kinetics; Membrane Glycoproteins; Membrane Lipids; Rhodamines; Temperature