B16 melanoma cell lines

Taniguchi, S., Eujiki, H., Kobayashi, H., et al.. Effect of (-)-epigallocatechin-3- gallate on lung metastasis with mouse B16 melanoma cell lines. Cancer Lett., 65, 51-54, 1992. 

Fig. 6.1. Selection of organ-specific variants with the B16 melanoma cell line. (A) B16-F10 were selected after repeated injections in the tail vein, and subcultivation of lung colonies. B16-LS9 have been selected by means of serial injections through the mesenteric veins, and subcultivation of liver colonies. (B) Livers of C57 Bl/6 mice taken 2 weeks after intrasplenic injection of B16-LS9 (two upper rows), or B16-F10 (two lower rows) cells. The superior liver colonization ability of B16-LS9 cells is evident also when the liver is the first capillary bed along the route of intravenously injected cells.

This method has been set up by using the B16 mouse melanoma cell line. A typical experiment is illustrated in Fig. 2.1(A, B). The application of this method to other cell lines might require little adjustments of some parameters, such as the time of incubation in PBS-CMF to detach the cells from the substrate as aggregates, and the shearing force to apply to reach a plateau value before trypsin addition. 

Cell filterability is influenced by a variety of biological and technological factors (Nordt, 1983). Thus, in order to be reliable and reproducible, Nucleopore filtration techniques must fulfil certain criteria, and namely (1) the most part of the input cells must be recovered in the filtrate (2) cellular aggregation should be minimized by choosing conditions which permit relatively short filtration times (3) cell viability should be high and not lost on filtration (4) cell size distribution should not be influenced by filtration and (5) differences in cell to filter and cell to cell adhesion of different cell lines should not be responsible for differences in filterability. In order to fulfil these criteria, experimental parameters such as cell to pore ratio, filtration pressure and cell culmre conditions have to be standardized. The following optimal conditions have been established for filtration of B16 melanoma cells (mean cell diameter 17.4 0.21 /xm, mean diameter of cell nuclei 9.8 0.27/xm) 20 cm H2O driving pressure, cell-to-pore ratio 1 1, temperature 22°C (Ochalek et al., 1988). Care has to be taken to derive tumor cells from similar culture conditions, since cell density has been found to influence filterability. 

A simple variation on the method described above was used by Tullberg et al. (1989) to select B16 melanoma cells of increased invasive (and metastatic) ability. Filters with pores of 10 /.tm are employed to avoid a selection for pore penetration, and coated with a thick layer (approximately 1 fim) of Matrigel , a reconstituted BM derived from EHS tumors (see Chapter 4 for more details). Cells are plated on the coated filters at a density that is lower than that used for the migration selection, since in this case the time it takes for the cells to migrate can be considerably higher, and excessive growth on the upper side of the filter has to be prevented. Optimal conditions have to be empirically determined for each cell line. Cells that have invaded and passed to the underside of the filter are collected and expanded as described above, and further selections can be made to obtain a fairly homogenous cell population of increased invasive potential, from which clones can be derived. 

ED50. zng/mL. J1D90 (ng/mL). MIC (ng/mL). 3L517y cell line, adenocarcinoma cell line. 7LoVo cell line. 8 B16 melanoma cells, drug-sensitive breast-cancer cell line. °HT-29 colon 9HeLa cells. I0T-47D ... 

The molecular mass of chitosan has an effect on the luciferase activities for A549 cells, B16 melanoma cells and Hela cells. Chitosans of 15 and 52 kDa largely promoted luciferase activity for all cell lines employed. The heptamer (1.3 kDa) did not show any transfection efficiency. The transfection efficiency mediated by 100 kDa chitosan was less than that of 15 and 52 kDa chitosans. Xue et al. synthesized carbohydrate-based polycations of a... 

The use of liposomes as boron delivery vehicles for BNCT has been extensively studied, since liposomes can transport large amounts of boronated molecules, both hydrophobic and hydrophilic in nature, selectively into tumor cells. Three methods for incorporation of boron into liposomes have been reported incorporation of hydrophobic molecules into lipid bilayers, entrapment of hydrophilic molecules into the liposomes aqueous core, and the use of boronated lipids. Recently, PEG-based liposomes (DPPC/cholesterol/DSPC-PEG2000) have been used as effective carriers of boronated compounds within tumor tissues. Intraveuously injected B-PEG-liposomes on human pancreatic carcinoma xenografts in nude mice inhibited tumor growth after thermal neutron irradiation. The cationic liposome formulation (DOTAP/DOPC/DOPE) was used to deliver ortho-carboranyl P-lactoside (LCOB) aud l-methyl-ort/j< -carboranyl-2-hexylthioporphyrazine (H2PZ-COB) within DHD/K12/TRb (DHD) rat colon carcinoma and B16-F10 murine melanoma cell lines. Results from these studies show that a thirty-fold increase on the cellular uptake of B was obtained for the boronated compounds in the liposome formulation as compared with that of the BPA-fructose complex. 

What started in the beginning of the century with the observation of cells taken from different sponges to allow the study of recognition processes [48, 49] has developed into a major research tool in biology with the use of both, primary and immortalized cells from many different sources. The selection of different clones from the same parent line has further stimulated the application and direct comparison of cells with similar but not identical surface composition. Examples of such cell lines that have been used for the study of carbohydrate-carbohydrate mediated phenomena are variants of the mouse melanoma cell line B16 expressing different levels of GM3-ceramides [79] or T-cell lymphoma clones with different levels of Gg3-ceramides on the cell surface [76] (see also Table 4). 

The cytotoxicity of diploicin (77) was evaluated by Millot and coworkers in 2009 against B16 (murine melanoma) and HaCaT (human keratinocyte) cell lines [100]. Pannarin (66), a secondary metabolite from lichen Diploicia canescens, was tested by two different research groups for its inhibitory potential against DU-145 prostate carcinoma [100] and M14 (human melanoma) cell lines with positive results [96]. Recently, the cytotoxicity of proto-lichesterinic acid (8) against HeLa cell lines has been evaluated by Brisdelli and coworkers. Its activity is based on its abdity to induce cell death through activation of caspases-3, -8, and -9 [101]. 

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