Abstract:Some of the greatest challenges in stem cells (SCs) biology and regenerative medicine are differentiation control of SCs and ensuring the purity of differentiated cells. In this work, we differentiated mouse pluripotent stem cells (mPSCs) toward pancreatic cells characterizing this differentiation process by molecular and spectroscopic technics. Both mPSCs and Differentiated Pancreatic Cells (DPCs) were subjected to a genetic, phenotypic, and biochemical analysis by real-time quantitative PCR (RT-qPCR), immuno… Show more
“…The present work aimed to evaluate the impact of EGCG consumption on the plasma molecular profile as obtained from high-throughput Fourier transform infrared (FTIR) spectroscopy, a very sensitive and specific technique. Examples of the technique's high sensitivity can be observed in applications to discriminate similar cells as bacteria serogroups from Salmonella [17], stem cell differentiation [18], cancer diagnosis and cancer grading [19,20], and to conduct diagnosis and prognosis of a diverse range of other diseases and the monitoring of treatments efficiency [21][22][23]. The high versatility of this technique also enables its application to a high diversity of biological samples, from tissue-based biopsies to body fluids such as urine, saliva, and tears [23][24][25].…”
Epigallocatechin-3-gallate (EGCG), the major catechin present in green tea, presents diverse appealing biological activities, such as antioxidative, anti-inflammatory, antimicrobial, and antiviral activities, among others. The present work evaluated the impact in the molecular profile of human plasma from daily consumption of 225 mg of EGCG for 90 days. Plasma from peripheral blood was collected from 30 healthy human volunteers and analyzed by high-throughput Fourier transform infrared spectroscopy. To capture the biochemical information while minimizing the interference of physical phenomena, several combinations of spectra pre-processing methods were evaluated by principal component analysis. The pre-processing method that led to the best class separation, that is, between the plasma spectral data collected at the beginning and after the 90 days, was a combination of atmospheric correction with a second derivative spectra. A hierarchical cluster analysis of second derivative spectra also highlighted the fact that plasma acquired before EGCG consumption presented a distinct molecular profile after the 90 days of EGCG consumption. It was also possible by partial least squares regression discriminant analysis to correctly predict all unlabeled plasma samples (not used for model construction) at both timeframes. We observed that the similarity in composition among the plasma samples was higher in samples collected after EGCG consumption when compared with the samples taken prior to EGCG consumption. Diverse negative peaks of the normalized second derivative spectra, associated with lipid and protein regions, were significantly affected (p < 0.001) by EGCG consumption, according to the impact of EGCG consumption on the patients' blood, low density and high density lipoproteins ratio. In conclusion, a single bolus dose of 225 mg of EGCG, ingested throughout a period of 90 days, drastically affected plasma molecular composition in all participants, which raises awareness regarding prolonged human exposure to EGCG. Because the analysis was conducted in a high-throughput, label-free, and economic analysis, it could be applied to high-dimension molecular epidemiological studies to further promote the understanding of the effect of bio-compound consumption mode and frequency.
“…The present work aimed to evaluate the impact of EGCG consumption on the plasma molecular profile as obtained from high-throughput Fourier transform infrared (FTIR) spectroscopy, a very sensitive and specific technique. Examples of the technique's high sensitivity can be observed in applications to discriminate similar cells as bacteria serogroups from Salmonella [17], stem cell differentiation [18], cancer diagnosis and cancer grading [19,20], and to conduct diagnosis and prognosis of a diverse range of other diseases and the monitoring of treatments efficiency [21][22][23]. The high versatility of this technique also enables its application to a high diversity of biological samples, from tissue-based biopsies to body fluids such as urine, saliva, and tears [23][24][25].…”
Epigallocatechin-3-gallate (EGCG), the major catechin present in green tea, presents diverse appealing biological activities, such as antioxidative, anti-inflammatory, antimicrobial, and antiviral activities, among others. The present work evaluated the impact in the molecular profile of human plasma from daily consumption of 225 mg of EGCG for 90 days. Plasma from peripheral blood was collected from 30 healthy human volunteers and analyzed by high-throughput Fourier transform infrared spectroscopy. To capture the biochemical information while minimizing the interference of physical phenomena, several combinations of spectra pre-processing methods were evaluated by principal component analysis. The pre-processing method that led to the best class separation, that is, between the plasma spectral data collected at the beginning and after the 90 days, was a combination of atmospheric correction with a second derivative spectra. A hierarchical cluster analysis of second derivative spectra also highlighted the fact that plasma acquired before EGCG consumption presented a distinct molecular profile after the 90 days of EGCG consumption. It was also possible by partial least squares regression discriminant analysis to correctly predict all unlabeled plasma samples (not used for model construction) at both timeframes. We observed that the similarity in composition among the plasma samples was higher in samples collected after EGCG consumption when compared with the samples taken prior to EGCG consumption. Diverse negative peaks of the normalized second derivative spectra, associated with lipid and protein regions, were significantly affected (p < 0.001) by EGCG consumption, according to the impact of EGCG consumption on the patients' blood, low density and high density lipoproteins ratio. In conclusion, a single bolus dose of 225 mg of EGCG, ingested throughout a period of 90 days, drastically affected plasma molecular composition in all participants, which raises awareness regarding prolonged human exposure to EGCG. Because the analysis was conducted in a high-throughput, label-free, and economic analysis, it could be applied to high-dimension molecular epidemiological studies to further promote the understanding of the effect of bio-compound consumption mode and frequency.
“…Other bands at 1154, 1055, and 1015 cm −1 respectively are related to C–O vibrations from glycogen and other carbohydrates [20]. Finally, a band around 991 cm −1 is mainly due to C–O stretch from RNA ribose chain [17, 23]. Fig.…”
Section: Resultsmentioning
confidence: 99%
“…On the other hand, the second derivative spectra of the same samples in the region of the amide I group (1700–1600 cm −1 ) calculated with Savitzky-Golay algorithm allowed the identification of various secondary structures present in the proteins, as β-turns (1694 and 1680 cm −1 ), α-helices (1650 cm −1 ), and β-pleated sheets (1633 cm −1 ) [17, 21–23]; in the same way, differences between mPSCs and DKCs on three bands at 1675, 1670, and 1633 cm −1 were observed, which are also related to asparagine, glutamine, and arginine respectively (Fig. 6b) [22].…”
BackgroundKidney diseases are a global health problem. Currently, over 2 million people require dialysis or transplant which are associated with high morbidity and mortality; therefore, new researches focused on regenerative medicine have been developed, including the use of stem cells.ResultsIn this research, we generate differentiated kidney cells (DKCs) from mouse pluripotent stem cells (mPSCs) analyzing their morphological, genetic, phenotypic, and spectroscopic characteristics along differentiation, highlighting that there are no reports of the use of Fourier transform infrared (FTIR) spectroscopy to characterize the directed differentiation of mPSCs to DKCs. The genetic and protein experiments proved the obtention of DKCs that passed through the chronological stages of embryonic kidney development. Regarding vibrational spectroscopy analysis by FTIR, bands related with biomolecules were shown on mPSCs and DKCs spectra, observing distinct differences between cell lineages and maturation stages. The second derivative of DKCs spectra showed changes in the protein bands compared to mPSCs. Finally, the principal components analysis obtained from FTIR spectra allowed to characterize chemical and structurally mPSCs and their differentiation process to DKCs in a rapid and non-invasive way.ConclusionOur results indicated that we obtained DKCs from mPSCs, which passed through the chronological stages of embryonic kidney development. Moreover, FTIR spectroscopy resulted in a non-invasive, rapid and precise technic that together with principal component analysis allows to characterize chemical and structurally both kind of cells and also discriminate and determine different stages along the cell differentiation process.
“…Realizando el análisis multivariado de ACP (Figura 5), pudimos observar en el primer componente principal para las tres regiones exploradas, una clara discrimi- nación entre CMP y CPD, evidenciando el cambio bioquímico y estructural que sufren las CM al ser diferenciadas, resultados que han sido reportados por diversos autores y nuestro grupo de investigación mediante técnicas moleculares o fenotípicas durante la diferenciación dirigida de CMP [2,4,5,8,13,14] .…”
Section: Tratamiento Espectral Y Análisis Multivariadounclassified
RESUMENDos de los grandes retos en la biología de las Células Madre (CM) y la Medicina Regenerativa, son el control en la diferenciación de estas células y asegurar la pureza de las células diferenciadas, por lo que es necesario contar con técnicas rápidas, eficientes y precisas para la caracterización de CM y su diferenciación a diferentes linajes celulares. El objetivo de este trabajo fue analizar Células Madre Pluripotentes (CMP) y Células Pancreáticas Diferenciadas (CPD) mediante espectroscopía Infrarroja por Transformada de Fourier (FTIR) y Análisis de Componentes Principales (ACP). Para ello se diferenciaron CMP a CPD, caracterizando el proceso de diferenciación a los días 0, 11, 17 y 21 mediante microscopía óptica y espectroscopia vibracional. Los espectros FTIR se analizaron con el método multivariado de ACP, utilizando su segunda derivada en las regiones de proteínas, carbohidratos y ribosas. Los resultados indican que el ACP permite caracterizar y discriminar CMP y CPD en sus diferentes etapas de diferenciación en las regiones espectrales analizadas. Con lo anterior concluimos que el ACP permite caracterizar química y estructuralmente CMP y diferentes etapas de su diferenciación en una forma rápida, precisa y no invasiva.
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