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

Abstract

BackgroundOne 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.MethodsHigh 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.ResultsSERS 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.ConclusionMolecular fingerprint and proliferation behavior of four B16 melanoma cell sub-lines were elucidated by associating SERS investigations with MTT proliferation assay.

Surface enhanced Raman spectroscopy measurements of MCF7 cells adhesion in confined micro-environments

“…In spectrum p2m1 we find the peaks at the 1232 cm À 1 , indicating the presence of the amide III band of proteins, the peak at 1350 cm À 1 assignable to adenine, guanine (DNA) [67], 1488 cm À 1 , stretching of purine [67] and the peaks at around 1570 cm À 1 , amide II band of proteins and at 1620 cm À 1 , amide I vibration. Also in spectrum p3m1 we find other peaks characteristic of nucleic cell part, in particular to nucleotides: 1574 cm À 1 , adenine and guanine, 1540 cm À 1 adenine [67] and 1394 cm À 1 [64].…”

“…In the spectrum p1m2 a strong enhancement is visible for some bands, at 1220 cm À 1 , which may be assigned to the OH deformation [61], at 1450 cm À 1 , assigned to the protein CH deformations vibration in deoxyribose [62], and at 1620 cm À 1 , amide I vibration. This point of the SERS Raman map feels principally the nucleus influence, as also the point p2m2 of the same map where the principal peaks are at 1230 cm À 1 , indicating membrane phospholipids [63], a weak peak around 1282 cm À 1 , CH bending vibrations [64], and the band around 1509 cm À 1 , assigned to guanine and adenine nucleic acid bases [64,65]. The same cell at point p3m2 shows the peak at 1225 cm À 1 , relative to the membrane phospholipids, a little shifted [63], the peaks at 1370, 1580 and 1628 cm À 1 evidencing the presence of cytochrome [66], and the band at 1656 cm À 1 of amide III.…”

“…The spectrum p1m1 marks the peaks at 870 cm À 1 , deoxyribose, at 1232 cm À 1 , at 1260 cm À 1 [67], DNA, at 1412 cm À 1 , assigned to amino acids and COO-asymmetric stretching vibrations [64], at 1509 cm À 1 , vibration of imidazole ring relative to nucleic acid bases, and the band at 1676 cm À 1 assigned to amide III β sheet. In spectrum p2m1 we find the peaks at the 1232 cm À 1 , indicating the presence of the amide III band of proteins, the peak at 1350 cm À 1 assignable to adenine, guanine (DNA) [67], 1488 cm À 1 , stretching of purine [67] and the peaks at around 1570 cm À 1 , amide II band of proteins and at 1620 cm À 1 , amide I vibration.…”

Considerations for the Uptake Characteristic of Inorganic Nanoparticles into Mammalian Cells—Insights Gained by TEM Investigations

“…In particular, transmission electron microscopy (TEM) investigations provide attractive possibilities to study the interaction of particles with biological barriers, i.e., the extracellular membrane or the nuclear membrane, their accumulation into cellular organelles or their spatial distribution on the scale of individual nanoparticles (e.g., uptake as individual nanoparticles or in form of large clusters, etc.). In addition TEM can simultaneously image also the cellular membranes and organelles to determine respective morphological changes upon nanoparticle interaction . Thus, attractive possibilities to acquire additional information on the particle fate inside the cell emerge from TEM investigations.…”

Imaging of red-shifted photons from bioluminescent tumours using fluorescence by unbound excitation from luminescence

“…In this study, we used two different in vivo subcutaneous bioluminescent tumour models 93 to investigate the suitability of FUEL in detecting tumours. The first model was induced 94 by bioluminescent B16-F10 tumour cells expressing firefly luciferase [19][20][21]. These 95 cells will be referred here as B16-Luc2.…”