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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

Time dependence SERS spectra acquired from a B164A5 cell. Excitation 633 nm. Laser power 5 mW.
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Figure 2: Time dependence SERS spectra acquired from a B164A5 cell. Excitation 633 nm. Laser power 5 mW.

Mentions: The SERS signal collected from different points across a cell exhibited a specific spectral shape, consistent with the nanoparticles accumulation. An enhanced Raman signal was obtained from the molecular species in the close vicinity of the nanoparticles. Since the laser spot encompasses an area of signal collection which depends on the objective magnification (i.e. 50x, NA 0.45), both regions, with and without nanoparticles, can fall in the laser spot and scatter the radiation. However, an interesting phenomenon was observed even for the signal acquired from the same focal spot, upon multiple accumulations. Such changes are illustrated in Figure 2 for a B164A5 cell. (Figure 2 Time dependence SERS spectra acquired from a B164A5 cell. Excitation 633 nm. Laser power 5 mW) The SERS spectra show intense bursts of Stokes shifted photons in an “on and off” manner exhibiting considerable fluctuations in both signal intensities and frequencies, over the course of the acquisition of 4 spectra in a total of 40 s. This phenomenon known as "blinking SERS" has been observed by different research groups [29,30]. The sudden appearance and disappearance of the SERS spectra was evidenced mainly for single molecule SERS experiments [8].


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)

Time dependence SERS spectra acquired from a B164A5 cell. Excitation 633 nm. Laser power 5 mW.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: Time dependence SERS spectra acquired from a B164A5 cell. Excitation 633 nm. Laser power 5 mW.
Mentions: The SERS signal collected from different points across a cell exhibited a specific spectral shape, consistent with the nanoparticles accumulation. An enhanced Raman signal was obtained from the molecular species in the close vicinity of the nanoparticles. Since the laser spot encompasses an area of signal collection which depends on the objective magnification (i.e. 50x, NA 0.45), both regions, with and without nanoparticles, can fall in the laser spot and scatter the radiation. However, an interesting phenomenon was observed even for the signal acquired from the same focal spot, upon multiple accumulations. Such changes are illustrated in Figure 2 for a B164A5 cell. (Figure 2 Time dependence SERS spectra acquired from a B164A5 cell. Excitation 633 nm. Laser power 5 mW) The SERS spectra show intense bursts of Stokes shifted photons in an “on and off” manner exhibiting considerable fluctuations in both signal intensities and frequencies, over the course of the acquisition of 4 spectra in a total of 40 s. This phenomenon known as "blinking SERS" has been observed by different research groups [29,30]. The sudden appearance and disappearance of the SERS spectra was evidenced mainly for single molecule SERS experiments [8].

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