A simple cell labeling technique by means of lectins linked to fluorochromes for the detection of cells on tissue sections.
We designed a protocol for cell labeling with the lectin wheat germ agglutinin (WGA) linked to the fluorochrome tetramethyl-rhodamine isothiocyanate (TRITC) for effective detection of the B16F10 melanoma and Lewis lung carcinoma (LLc) cells on pulmonary histological sections from C57BL/6 mice. We have also determined a suitable concentration of WGA-TRITC (10 micrograms/ml), which leads to a very intense and homogeneous labeling of the cells, as it avoids cell clumping due to the presence of the lectin WGA. In order to determine to what extent the method affects these tumor cells, we have studied some important aspects related to their metastatic behavior, taking into account three parameters: a) viability and rate of proliferation of the cells cultured in vitro; b) percentage of animals (C57BL/6 mice) bearing metastasis 15 days after intravenous inoculation with 10(5) B16F10 or LLc cells; and c) pattern of distribution of tumor foci in lung. There were no significant differences in these three parameters between the WGA-TRITC labeled-cells compared to the cultures of non-labeled cells in either of the cell lines (B16F10, LLc). Thus, we conclude that B16F10 and LLc tumor cells can be labeled following the protocol set-up in our study, as it allows these cells to be neatly identified on tissue sections and it causes no important physiological changes in the cells, with regard to metastatic behavior. These points make this technique very suitable for the detection of B16F10 and LLc cells on histological sections in studying their behavior during the first stages of the metastatic process.
Alonso-Varona
A
A
Dpto Biología Celular y Ciencias Morfológicas, Fac Medicina y Odontología, Univ del País Vasco-Euskal Herriko Unibertsitatea, Barrio Sarriena sn Leioa, Vizcaya, Spain.
Calle
Y
Y
Palomares
T
T
Castro
B
B
Barberá-Guillem
E
E
D000818:Animals; D018827:Carcinoma, Lewis Lung / Q000473:pathology* / Q000556:secondary; D002455:Cell Division; D002470:Cell Survival; D003584:Cytological Techniques; D005260:Female; D005456:Fluorescent Dyes; D008545:Melanoma / Q000473:pathology* / Q000556:secondary; D051379:Mice; D008810:Mice, Inbred C57BL; D012235:Rhodamines; D014407:Tumor Cells, Cultured; D014909:Wheat Germ Agglutinins
D005456:Fluorescent Dyes; D012235:Rhodamines; D014909:Wheat Germ Agglutinins; C009434:tetramethylrhodamine isothiocyanate