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A characterization of four B16 murine melanoma cell sublines molecular fingerprint and proliferation behavior.

Danciu C, Falamas A, Dehelean C, Soica C, Radeke H, Barbu-Tudoran L, Bojin F, Pînzaru SC, Munteanu MF - Cancer Cell Int. (2013)

Bottom Line: SERS bands allowed the identification inside the cells of the main bio-molecular components such as: proteins, nucleic acids, and lipids.An "on and off" SERS effect was constantly present, which may be explained in terms of the employed laser power, as well as the possible different orientations of the adsorbed species in the cells in respect to the Ag nanoparticles.MTT results showed that among the four tested cell sub-lines B16 F10 is the most proliferative and B164A5 has the lower growth capacity.

View Article: PubMed Central - HTML - PubMed

Affiliation: Faculty of Pharmacy, University of Medicine and Pharmacy "Victor Babes", EftimieMurgu Square, No. 2, 300041 Timişoara, România.

ABSTRACT

Background: One of the most popular and versatile model of murine melanoma is by inoculating B16 cells in the syngeneic C57BL6J mouse strain. A characterization of different B16 modified cell sub-lines will be of real practical interest. For this aim, modern analytical tools like surface enhanced Raman spectroscopy/scattering (SERS) and MTT were employed to characterize both chemical composition and proliferation behavior of the selected cells.

Methods: High quality SERS signal was recorded from each of the four types of B16 cell sub-lines: B164A5, B16GMCSF, B16FLT3, B16F10, in order to observe the differences between a parent cell line (B164A5) and other derived B16 cell sub-lines. Cells were incubated with silver nanoparticles of 50-100 nm diameter and the nanoparticles uptake inside the cells cytoplasm was proved by transmission electron microscopy (TEM) investigations. In order to characterize proliferation, growth curves of the four B16 cell lines, using different cell numbers and FCS concentration were obtained employing the MTT proliferation assay. For correlations doubling time were calculated.

Results: SERS bands allowed the identification inside the cells of the main bio-molecular components such as: proteins, nucleic acids, and lipids. An "on and off" SERS effect was constantly present, which may be explained in terms of the employed laser power, as well as the possible different orientations of the adsorbed species in the cells in respect to the Ag nanoparticles. MTT results showed that among the four tested cell sub-lines B16 F10 is the most proliferative and B164A5 has the lower growth capacity. Regarding B16FLT3 cells and B16GMCSF cells, they present proliferation ability in between with slight slower potency for B16GMCSF cells.

Conclusion: Molecular fingerprint and proliferation behavior of four B16 melanoma cell sub-lines were elucidated by associating SERS investigations with MTT proliferation assay.

No MeSH data available.


Related in: MedlinePlus

Doubling time of the four B16 cell sublines as revealed by trypan blue assay.
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Figure 6: Doubling time of the four B16 cell sublines as revealed by trypan blue assay.

Mentions: Assuming a constant growth rate (increase per unit of time is proportional to the current quantity), the results showed that B164A5 cells present the longest doubling time, 24 h. The lowest value vas obtained for B16F10 cells, 17.2 h. For the transduced B16GMCSF cells doubling time was 17.9 h and for the transfected B16FLT3 cells the value was 18.4 h. The first conclusion after this assay is that B16F10 cells have the smallest doubling time so the biggest multiplication ratio while B164A5 cells have the biggest doubling time so the smallest multiplication ratio. B16GMCSF cells and B16FLT3 cells present values in between, with a slight higher value in the case of B16FLT3 cells. Results can be seen in Figure 6 Values obtained for the doubling time of the four cell lines. In another study Ohira et al. found the doubling time of B16F10 cells 20.1 h, while Yerlikaya et al. found the doubling time of B16F10 cells 14.2 h [38,39]. Probably variable values are found by different research groups because of different chosen compositions of the growing medium. Qarawi et al. estimated that the doubling time for B16 cells is approximately 24 h [40].


A characterization of four B16 murine melanoma cell sublines molecular fingerprint and proliferation behavior.

Danciu C, Falamas A, Dehelean C, Soica C, Radeke H, Barbu-Tudoran L, Bojin F, Pînzaru SC, Munteanu MF - Cancer Cell Int. (2013)

Doubling time of the four B16 cell sublines as revealed by trypan blue assay.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750233&req=5

Figure 6: Doubling time of the four B16 cell sublines as revealed by trypan blue assay.
Mentions: Assuming a constant growth rate (increase per unit of time is proportional to the current quantity), the results showed that B164A5 cells present the longest doubling time, 24 h. The lowest value vas obtained for B16F10 cells, 17.2 h. For the transduced B16GMCSF cells doubling time was 17.9 h and for the transfected B16FLT3 cells the value was 18.4 h. The first conclusion after this assay is that B16F10 cells have the smallest doubling time so the biggest multiplication ratio while B164A5 cells have the biggest doubling time so the smallest multiplication ratio. B16GMCSF cells and B16FLT3 cells present values in between, with a slight higher value in the case of B16FLT3 cells. Results can be seen in Figure 6 Values obtained for the doubling time of the four cell lines. In another study Ohira et al. found the doubling time of B16F10 cells 20.1 h, while Yerlikaya et al. found the doubling time of B16F10 cells 14.2 h [38,39]. Probably variable values are found by different research groups because of different chosen compositions of the growing medium. Qarawi et al. estimated that the doubling time for B16 cells is approximately 24 h [40].

Bottom Line: SERS bands allowed the identification inside the cells of the main bio-molecular components such as: proteins, nucleic acids, and lipids.An "on and off" SERS effect was constantly present, which may be explained in terms of the employed laser power, as well as the possible different orientations of the adsorbed species in the cells in respect to the Ag nanoparticles.MTT results showed that among the four tested cell sub-lines B16 F10 is the most proliferative and B164A5 has the lower growth capacity.

View Article: PubMed Central - HTML - PubMed

Affiliation: Faculty of Pharmacy, University of Medicine and Pharmacy "Victor Babes", EftimieMurgu Square, No. 2, 300041 Timişoara, România.

ABSTRACT

Background: One of the most popular and versatile model of murine melanoma is by inoculating B16 cells in the syngeneic C57BL6J mouse strain. A characterization of different B16 modified cell sub-lines will be of real practical interest. For this aim, modern analytical tools like surface enhanced Raman spectroscopy/scattering (SERS) and MTT were employed to characterize both chemical composition and proliferation behavior of the selected cells.

Methods: High quality SERS signal was recorded from each of the four types of B16 cell sub-lines: B164A5, B16GMCSF, B16FLT3, B16F10, in order to observe the differences between a parent cell line (B164A5) and other derived B16 cell sub-lines. Cells were incubated with silver nanoparticles of 50-100 nm diameter and the nanoparticles uptake inside the cells cytoplasm was proved by transmission electron microscopy (TEM) investigations. In order to characterize proliferation, growth curves of the four B16 cell lines, using different cell numbers and FCS concentration were obtained employing the MTT proliferation assay. For correlations doubling time were calculated.

Results: SERS bands allowed the identification inside the cells of the main bio-molecular components such as: proteins, nucleic acids, and lipids. An "on and off" SERS effect was constantly present, which may be explained in terms of the employed laser power, as well as the possible different orientations of the adsorbed species in the cells in respect to the Ag nanoparticles. MTT results showed that among the four tested cell sub-lines B16 F10 is the most proliferative and B164A5 has the lower growth capacity. Regarding B16FLT3 cells and B16GMCSF cells, they present proliferation ability in between with slight slower potency for B16GMCSF cells.

Conclusion: Molecular fingerprint and proliferation behavior of four B16 melanoma cell sub-lines were elucidated by associating SERS investigations with MTT proliferation assay.

No MeSH data available.


Related in: MedlinePlus