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Tracking adipogenic differentiation of skeletal stem cells by label-free chemically selective imaging

Tracking adipogenic differentiation of skeletal stem cells by label-free chemically selective imaging
Tracking adipogenic differentiation of skeletal stem cells by label-free chemically selective imaging
Coherent anti-Stokes Raman scattering (CARS) is a chemically selective label-free imaging technique which is rapidly emerging as a powerful alternative to conventional microscopy in biomedicine. The strength of this imaging approach is the provision of rapid insight into chemical distribution especially of small biomolecules such as lipids. The label-free, non-destructive and non-invasive nature of CARS lends itself for use with stem cells, as labelling or staining will render them otherwise unsuitable for therapy. Isolation, enrichment and characterisation of skeletal stem cells (SSCs) and their progeny is of tremendous significance in regenerative medicine. However, SSCs differentiation into bone, cartilage or fat cell types, is currently assessed using several invasive and, typically, destructive methodologies. Thus, CARS presents an exciting alternative to interrogate the differentiation of SSCs in their natural state. In the current study, we have examined the adipogenic differentiation of SSCs over time using CARS imaging and verified the observed differentiation using molecular analysis of gene expression as well as compared the results to conventional Oil Red O lipid staining. We find that the CARS analysis provides an enhanced resolution and definition of lipid droplets, detectable as early as 24 hours and 72 hours after adipogenic induction. Quantification of the CARS image data sets also showed a change in lipid droplet size distribution during the course of adipogenesis over 14 days. Furthermore, CARS provided a superior and facile approach to monitor changes in SSCs as a result of chemical modulation of adipogenic differentiation. The current studies pave the way for the use of CARS as a powerful chemical imaging tool in therapeutics, regenerative medicine and skeletal stem cell biology
1478-6524
7089-7096
Smus, Justyna P.
dd94b2b9-f0bc-4221-8ca8-40265463d9f6
Moura, Catarina Costa
75550295-bd74-4dad-b0fc-75c02fdbe54d
McMorrow, Emma
448100b1-cdf7-4530-9431-8dbdfd591132
Tare, Rahul S.
587c9db4-e409-4e7c-a02a-677547ab724a
Oreffo, Richard O.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Mahajan, Sumeet
b131f40a-479e-4432-b662-19d60d4069e9
Smus, Justyna P.
dd94b2b9-f0bc-4221-8ca8-40265463d9f6
Moura, Catarina Costa
75550295-bd74-4dad-b0fc-75c02fdbe54d
McMorrow, Emma
448100b1-cdf7-4530-9431-8dbdfd591132
Tare, Rahul S.
587c9db4-e409-4e7c-a02a-677547ab724a
Oreffo, Richard O.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Mahajan, Sumeet
b131f40a-479e-4432-b662-19d60d4069e9

Smus, Justyna P., Moura, Catarina Costa, McMorrow, Emma, Tare, Rahul S., Oreffo, Richard O.C. and Mahajan, Sumeet (2015) Tracking adipogenic differentiation of skeletal stem cells by label-free chemically selective imaging. Chemical Science, 6 (12), 7089-7096. (doi:10.1039/C5SC02168E).

Record type: Article

Abstract

Coherent anti-Stokes Raman scattering (CARS) is a chemically selective label-free imaging technique which is rapidly emerging as a powerful alternative to conventional microscopy in biomedicine. The strength of this imaging approach is the provision of rapid insight into chemical distribution especially of small biomolecules such as lipids. The label-free, non-destructive and non-invasive nature of CARS lends itself for use with stem cells, as labelling or staining will render them otherwise unsuitable for therapy. Isolation, enrichment and characterisation of skeletal stem cells (SSCs) and their progeny is of tremendous significance in regenerative medicine. However, SSCs differentiation into bone, cartilage or fat cell types, is currently assessed using several invasive and, typically, destructive methodologies. Thus, CARS presents an exciting alternative to interrogate the differentiation of SSCs in their natural state. In the current study, we have examined the adipogenic differentiation of SSCs over time using CARS imaging and verified the observed differentiation using molecular analysis of gene expression as well as compared the results to conventional Oil Red O lipid staining. We find that the CARS analysis provides an enhanced resolution and definition of lipid droplets, detectable as early as 24 hours and 72 hours after adipogenic induction. Quantification of the CARS image data sets also showed a change in lipid droplet size distribution during the course of adipogenesis over 14 days. Furthermore, CARS provided a superior and facile approach to monitor changes in SSCs as a result of chemical modulation of adipogenic differentiation. The current studies pave the way for the use of CARS as a powerful chemical imaging tool in therapeutics, regenerative medicine and skeletal stem cell biology

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Accepted/In Press date: 8 September 2015
e-pub ahead of print date: 9 September 2015
Published date: 9 September 2015
Organisations: Institute for Life Sciences, Chemical Biology Group

Identifiers

Local EPrints ID: 382413
URI: http://eprints.soton.ac.uk/id/eprint/382413
ISSN: 1478-6524
PURE UUID: bdf7e54c-64f9-4fdb-93d5-e7ed97a75b6a
ORCID for Rahul S. Tare: ORCID iD orcid.org/0000-0001-8274-8837
ORCID for Richard O.C. Oreffo: ORCID iD orcid.org/0000-0001-5995-6726
ORCID for Sumeet Mahajan: ORCID iD orcid.org/0000-0001-8923-6666

Catalogue record

Date deposited: 30 Oct 2015 09:58
Last modified: 13 Nov 2021 02:45

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Contributors

Author: Justyna P. Smus
Author: Catarina Costa Moura
Author: Emma McMorrow
Author: Rahul S. Tare ORCID iD
Author: Sumeet Mahajan ORCID iD

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