Limits...
The characterisation of pluripotent and multipotent stem cells using Fourier transform infrared microspectroscopy.

Cao J, Ng ES, McNaughton D, Stanley EG, Elefanty AG, Tobin MJ, Heraud P - Int J Mol Sci (2013)

Bottom Line: Fourier transform infrared (FTIR) microspectroscopy shows potential as a benign, objective and rapid tool to screen pluripotent and multipotent stem cells for clinical use.It offers a new experimental approach that provides a holistic measurement of macromolecular composition such that a signature representing the internal cellular phenotype is obtained.The use of this technique therefore contributes information that is complementary to that acquired by conventional genetic and immunohistochemical methods.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Wellington Road, Clayton, Victoria 3800, Australia. phil.heraud@monash.edu.

ABSTRACT
Fourier transform infrared (FTIR) microspectroscopy shows potential as a benign, objective and rapid tool to screen pluripotent and multipotent stem cells for clinical use. It offers a new experimental approach that provides a holistic measurement of macromolecular composition such that a signature representing the internal cellular phenotype is obtained. The use of this technique therefore contributes information that is complementary to that acquired by conventional genetic and immunohistochemical methods.

Show MeSH

Related in: MedlinePlus

(A) Scores plots for PLS-DA of the spectral data for the three treatment groups from one experiment showing replicate samples for each cell type. Each spectrum is represented as a point in PC1 vs. PC2 vs. PC3 space; (B) Regression coefficient plots used to explain the clustering observed in the PLS scores plot shown in panel A. PC1 regression coefficient loadings indicated spectral differences explaining clustering along PC1, which separates hESCs from the cells differentiated in FGF2 or BMP4/Act A. PC3 regression coefficient loadings explained the separation of the spectra from FGF2-and BMP4/Act A-treated sample groups along PC3. Limited spread in the PC2 direction did not contribute significantly to the clustering of samples [32].
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC3794735&req=5

f5-ijms-14-17453: (A) Scores plots for PLS-DA of the spectral data for the three treatment groups from one experiment showing replicate samples for each cell type. Each spectrum is represented as a point in PC1 vs. PC2 vs. PC3 space; (B) Regression coefficient plots used to explain the clustering observed in the PLS scores plot shown in panel A. PC1 regression coefficient loadings indicated spectral differences explaining clustering along PC1, which separates hESCs from the cells differentiated in FGF2 or BMP4/Act A. PC3 regression coefficient loadings explained the separation of the spectra from FGF2-and BMP4/Act A-treated sample groups along PC3. Limited spread in the PC2 direction did not contribute significantly to the clustering of samples [32].

Mentions: In this work, FPA-FTIR microspectroscopy was used to acquire spectra of hESC that were differentiated towards mesendoderm (precursor cells of mesoderm and endodermal embryonic germ layers) induced by a bone morphogenetic protein cytokine cocktail (BMP4/Act A medium) or towards ectodermal lineages (precursor cells of the skin and nervous system) in medium supplemented with fibroblast growth factor (FGF2) (Figure 5). Across all three experimental replicates, the undifferentiated human embryonic stem cell line, MIXL1GFP/w, could be discriminated from its differentiated progeny due to spectra of the undifferentiated cells possessing higher IR absorbance for lipid and glycogen components. The distinctness of the spectra of the progeny cells from the spectra of the stem cells was further confirmed using various multivariate data analysis approaches including PCA, PLS-DA and ANN modelling.


The characterisation of pluripotent and multipotent stem cells using Fourier transform infrared microspectroscopy.

Cao J, Ng ES, McNaughton D, Stanley EG, Elefanty AG, Tobin MJ, Heraud P - Int J Mol Sci (2013)

(A) Scores plots for PLS-DA of the spectral data for the three treatment groups from one experiment showing replicate samples for each cell type. Each spectrum is represented as a point in PC1 vs. PC2 vs. PC3 space; (B) Regression coefficient plots used to explain the clustering observed in the PLS scores plot shown in panel A. PC1 regression coefficient loadings indicated spectral differences explaining clustering along PC1, which separates hESCs from the cells differentiated in FGF2 or BMP4/Act A. PC3 regression coefficient loadings explained the separation of the spectra from FGF2-and BMP4/Act A-treated sample groups along PC3. Limited spread in the PC2 direction did not contribute significantly to the clustering of samples [32].
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3794735&req=5

f5-ijms-14-17453: (A) Scores plots for PLS-DA of the spectral data for the three treatment groups from one experiment showing replicate samples for each cell type. Each spectrum is represented as a point in PC1 vs. PC2 vs. PC3 space; (B) Regression coefficient plots used to explain the clustering observed in the PLS scores plot shown in panel A. PC1 regression coefficient loadings indicated spectral differences explaining clustering along PC1, which separates hESCs from the cells differentiated in FGF2 or BMP4/Act A. PC3 regression coefficient loadings explained the separation of the spectra from FGF2-and BMP4/Act A-treated sample groups along PC3. Limited spread in the PC2 direction did not contribute significantly to the clustering of samples [32].
Mentions: In this work, FPA-FTIR microspectroscopy was used to acquire spectra of hESC that were differentiated towards mesendoderm (precursor cells of mesoderm and endodermal embryonic germ layers) induced by a bone morphogenetic protein cytokine cocktail (BMP4/Act A medium) or towards ectodermal lineages (precursor cells of the skin and nervous system) in medium supplemented with fibroblast growth factor (FGF2) (Figure 5). Across all three experimental replicates, the undifferentiated human embryonic stem cell line, MIXL1GFP/w, could be discriminated from its differentiated progeny due to spectra of the undifferentiated cells possessing higher IR absorbance for lipid and glycogen components. The distinctness of the spectra of the progeny cells from the spectra of the stem cells was further confirmed using various multivariate data analysis approaches including PCA, PLS-DA and ANN modelling.

Bottom Line: Fourier transform infrared (FTIR) microspectroscopy shows potential as a benign, objective and rapid tool to screen pluripotent and multipotent stem cells for clinical use.It offers a new experimental approach that provides a holistic measurement of macromolecular composition such that a signature representing the internal cellular phenotype is obtained.The use of this technique therefore contributes information that is complementary to that acquired by conventional genetic and immunohistochemical methods.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Developmental Biology, Faculty of Medicine, Nursing and Health Sciences, Monash University, Wellington Road, Clayton, Victoria 3800, Australia. phil.heraud@monash.edu.

ABSTRACT
Fourier transform infrared (FTIR) microspectroscopy shows potential as a benign, objective and rapid tool to screen pluripotent and multipotent stem cells for clinical use. It offers a new experimental approach that provides a holistic measurement of macromolecular composition such that a signature representing the internal cellular phenotype is obtained. The use of this technique therefore contributes information that is complementary to that acquired by conventional genetic and immunohistochemical methods.

Show MeSH
Related in: MedlinePlus