Discrimination of micromass-induced chondrocytes from human mesenchymal stem cells by focal plane array-Fourier transform infrared microspectroscopy

Chirapond Chonanant, Keith R Bambery, Nichada Jearanaikoon, Sirinart Chio-Srichan, Temduang Limpaiboon, Mark J Tobin, Phil R Heraud, Patcharee Jearanaikoon

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Rapid and sensitive methods for identifying stem cell differentiation state are required for facilitating future stem cell therapies. We aimed to evaluate the capability of focal plane array-Fourier transform infrared (FPA-FTIR) microspectroscopy for characterising the differentiation of chondrocytes from human mesenchymal stem cells (hMSCs). Successful induction was validated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis for collagen and aggrecan expression as chondrocyte markers in parallel with the spectroscopy. Spectra derived from chondrocyte-induced cells revealed strong IR absorbance bands attributed to collagen near 1338 and 1234cm(-1) and proteoglycan at 1245 and 1175-960cm(-1) compared to the non-induced cells. In addition, spectra from control and induced cells are segregated into separate clusters in partial least squares discriminant analysis score plots at the very early stages of induction and discrimination of an independent set of validation spectra with 100 accuracy. The predominant bands responsible for this discrimination were associated with collagen and aggrecan protein concordant with those obtained from RT-PCR and Western blot techniques. Our findings support the capability of FPA-FTIR microspectroscopy as a label-free tool for stem cell characterization allowing rapid and sensitive detection of macromolecular changes during chondrogenic differentiation.
Original languageEnglish
Pages (from-to)39 - 48
Number of pages10
JournalTalanta
Volume130
DOIs
Publication statusPublished - 2014

Cite this

Chonanant, C., Bambery, K. R., Jearanaikoon, N., Chio-Srichan, S., Limpaiboon, T., Tobin, M. J., ... Jearanaikoon, P. (2014). Discrimination of micromass-induced chondrocytes from human mesenchymal stem cells by focal plane array-Fourier transform infrared microspectroscopy. Talanta, 130, 39 - 48. https://doi.org/10.1016/j.talanta.2014.05.037
Chonanant, Chirapond ; Bambery, Keith R ; Jearanaikoon, Nichada ; Chio-Srichan, Sirinart ; Limpaiboon, Temduang ; Tobin, Mark J ; Heraud, Phil R ; Jearanaikoon, Patcharee. / Discrimination of micromass-induced chondrocytes from human mesenchymal stem cells by focal plane array-Fourier transform infrared microspectroscopy. In: Talanta. 2014 ; Vol. 130. pp. 39 - 48.
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title = "Discrimination of micromass-induced chondrocytes from human mesenchymal stem cells by focal plane array-Fourier transform infrared microspectroscopy",
abstract = "Rapid and sensitive methods for identifying stem cell differentiation state are required for facilitating future stem cell therapies. We aimed to evaluate the capability of focal plane array-Fourier transform infrared (FPA-FTIR) microspectroscopy for characterising the differentiation of chondrocytes from human mesenchymal stem cells (hMSCs). Successful induction was validated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis for collagen and aggrecan expression as chondrocyte markers in parallel with the spectroscopy. Spectra derived from chondrocyte-induced cells revealed strong IR absorbance bands attributed to collagen near 1338 and 1234cm(-1) and proteoglycan at 1245 and 1175-960cm(-1) compared to the non-induced cells. In addition, spectra from control and induced cells are segregated into separate clusters in partial least squares discriminant analysis score plots at the very early stages of induction and discrimination of an independent set of validation spectra with 100 accuracy. The predominant bands responsible for this discrimination were associated with collagen and aggrecan protein concordant with those obtained from RT-PCR and Western blot techniques. Our findings support the capability of FPA-FTIR microspectroscopy as a label-free tool for stem cell characterization allowing rapid and sensitive detection of macromolecular changes during chondrogenic differentiation.",
author = "Chirapond Chonanant and Bambery, {Keith R} and Nichada Jearanaikoon and Sirinart Chio-Srichan and Temduang Limpaiboon and Tobin, {Mark J} and Heraud, {Phil R} and Patcharee Jearanaikoon",
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Discrimination of micromass-induced chondrocytes from human mesenchymal stem cells by focal plane array-Fourier transform infrared microspectroscopy. / Chonanant, Chirapond; Bambery, Keith R; Jearanaikoon, Nichada; Chio-Srichan, Sirinart; Limpaiboon, Temduang; Tobin, Mark J; Heraud, Phil R; Jearanaikoon, Patcharee.

In: Talanta, Vol. 130, 2014, p. 39 - 48.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Discrimination of micromass-induced chondrocytes from human mesenchymal stem cells by focal plane array-Fourier transform infrared microspectroscopy

AU - Chonanant, Chirapond

AU - Bambery, Keith R

AU - Jearanaikoon, Nichada

AU - Chio-Srichan, Sirinart

AU - Limpaiboon, Temduang

AU - Tobin, Mark J

AU - Heraud, Phil R

AU - Jearanaikoon, Patcharee

PY - 2014

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N2 - Rapid and sensitive methods for identifying stem cell differentiation state are required for facilitating future stem cell therapies. We aimed to evaluate the capability of focal plane array-Fourier transform infrared (FPA-FTIR) microspectroscopy for characterising the differentiation of chondrocytes from human mesenchymal stem cells (hMSCs). Successful induction was validated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis for collagen and aggrecan expression as chondrocyte markers in parallel with the spectroscopy. Spectra derived from chondrocyte-induced cells revealed strong IR absorbance bands attributed to collagen near 1338 and 1234cm(-1) and proteoglycan at 1245 and 1175-960cm(-1) compared to the non-induced cells. In addition, spectra from control and induced cells are segregated into separate clusters in partial least squares discriminant analysis score plots at the very early stages of induction and discrimination of an independent set of validation spectra with 100 accuracy. The predominant bands responsible for this discrimination were associated with collagen and aggrecan protein concordant with those obtained from RT-PCR and Western blot techniques. Our findings support the capability of FPA-FTIR microspectroscopy as a label-free tool for stem cell characterization allowing rapid and sensitive detection of macromolecular changes during chondrogenic differentiation.

AB - Rapid and sensitive methods for identifying stem cell differentiation state are required for facilitating future stem cell therapies. We aimed to evaluate the capability of focal plane array-Fourier transform infrared (FPA-FTIR) microspectroscopy for characterising the differentiation of chondrocytes from human mesenchymal stem cells (hMSCs). Successful induction was validated by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis for collagen and aggrecan expression as chondrocyte markers in parallel with the spectroscopy. Spectra derived from chondrocyte-induced cells revealed strong IR absorbance bands attributed to collagen near 1338 and 1234cm(-1) and proteoglycan at 1245 and 1175-960cm(-1) compared to the non-induced cells. In addition, spectra from control and induced cells are segregated into separate clusters in partial least squares discriminant analysis score plots at the very early stages of induction and discrimination of an independent set of validation spectra with 100 accuracy. The predominant bands responsible for this discrimination were associated with collagen and aggrecan protein concordant with those obtained from RT-PCR and Western blot techniques. Our findings support the capability of FPA-FTIR microspectroscopy as a label-free tool for stem cell characterization allowing rapid and sensitive detection of macromolecular changes during chondrogenic differentiation.

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