FTIR Microspectroscopy for the Assessment of Mycoplasmas in HepG2 Cell Culture

Abstract

To assess the presence and absence of mycoplasma contamination in cell culture, Fourier transform infrared (FTIR) microspectroscopy, coupled with multivariate analysis, was deployed to determine the biomolecular changes in hepatocellular carcinoma cells, HepG2, before and after mycoplasma contamination. The contaminated HepG2 cells were treated with antibiotic BM-Cyclin to decontaminate the mycoplasma, and polymerase chain reaction (PCR) was then performed to confirm the presence or the absence of mycoplasma contamination. The contaminated and decontaminated HepG2 cells were analyzed by FTIR microspectroscopy with principal component analysis (PCA) and peak integral area analysis. The results showed that the FTIR spectra of contaminated HepG2 cells demonstrated the alteration in the IR spectra corresponding to the lipid, protein, and nucleic acid regions. PCA analysis distinguished the spectral differences between the groups of mycoplasma-contaminated and -decontaminated cells. The PCA loading plots suggest that lipid and protein are the main contributed molecules for the difference between these two cell groups. Peak integral area analysis illustrated the increase of lipid and nucleic acid and the decrease of protein contents in the contaminated HepG2 cells. FTIR microspectroscopy is, therefore, proven to be a potential tool for assessing mycoplasma removal by monitoring biomolecular alterations in cell culture.

Cytotoxicity, Removal of Congo Red Dye in Aqueous Solution Using Synthesized Amorphous Iron Oxide Nanoparticles from Incense Sticks Ash Waste

“…The band at 1630 cm -1 and a broad-band near 3400 cm -1 is assigned to the OH group of a H 2 O molecule/or Fe(OH) 3 present in the sample [47], whose results are in close agreement with Yadav et al, 2020 [48], where Fe-O bands were obtained in the range of 400-900 cm -1 , and water molecule band at 1660 and 3428 cm -1 [6,26]. A small band near 2800 cm -1 is could be due to the adsorption of atmospheric CO 2 by the sample as reported by numerous investigators in the literature [49]. 3.…”

AFM Study of Nanoscale Membrane Perturbation Induced by Antimicrobial Lipopeptide C14 KYR

“…Approximately at a given region of each condition, over 100 spectra were collected and analyzed. Figure 1 shows the representative second-derivative IR spectra in a certain range with assigned bands corresponding to the presence of macromolecules as follows: (A) lipid (3000–2800 cm −1 ), (B) protein (1700–1500 cm −1 ), and (C) carbohydrate and nucleic acid (1300–900 cm −1 ) [ 24 , 25 , 26 , 36 ]. When compared with the untreated cells, alterations in membrane components in all types of macromolecules were observed, as indicated by the differences in bands highlighted in yellow.…”

“…The main alteration in these regions of the MIC condition lightly shifted the peak, while 5xMIC C 14 -KYR addition was detectable with the decrease in the shift of the absorption peak, indicating the change in C-H stretching of the fatty-tail structure. The modification of the C-H vibration on the exposed bacterial profile is interpreted as bacterial lysis [ 26 , 34 , 36 ].…”

“…Then, cells were washed three times with 0.1% ( w / v ) NaCl and resuspended in 0.1% ( w / v ) NaCl. The cell suspension was transferred onto Low-e Microscope Slides (MirrIR, Kevley Technologies, Chesterland, OH, USA) and was vacuum-dried for 30 min in a desiccator [ 23 , 24 , 25 , 26 ]. The cells on the slide were rinsed with distilled water and then vacuum-dried.…”

New cross‐linked poly(methyl methacrylate): Synthesis, characterization, and inhibitory effects against selected bacteria and cancer cells

“…After the incubation period, microscopic images were taken using inverted microscopy then HepG2 cells were collected and counted using a hemocytometer. [ 24,25 ]…”