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  High content image analysis of focal adhesion-dependent mechanosensitive stem cell differentiation

Holle, A. W., McIntyre, A. J., Kehe, J., Wijesekara, P., Young, J. L., Vincent, L. G., et al. (2016). High content image analysis of focal adhesion-dependent mechanosensitive stem cell differentiation. Integrative Biology, 8(10), 1049-1058. doi:10.1039/C6IB00076B.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-A9E3-8 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002E-2F8F-2
Genre: Journal Article

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 Creators:
Holle, Andrew W., Author
McIntyre, Alistair J., Author
Kehe, Jared, Author
Wijesekara, Piyumi, Author
Young, Jennifer L.1, Author              
Vincent, Ludovic G., Author
Engler, Adam J., Author
Affiliations:
1Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society, ou_2364731              

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 Abstract: Human mesenchymal stem cells (hMSCs) receive differentiation cues from a number of stimuli, including extracellular matrix (ECM) stiffness. The pathways used to sense stiffness and other physical cues are just now being understood and include proteins within focal adhesions. To rapidly advance the pace of discovery for novel mechanosensitive proteins, we employed a combination of in silico and high throughput in vitro methods to analyze 47 different focal adhesion proteins for cryptic kinase binding sites. High content imaging of hMSCs treated with small interfering RNAs for the top 6 candidate proteins showed novel effects on both osteogenic and myogenic differentiation; Vinculin and SORBS1 were necessary for stiffness-mediated myogenic and osteogenic differentiation, respectively. Both of these proteins bound to MAPK1 (also known as ERK2), suggesting that it plays a context-specific role in mechanosensing for each lineage; validation for these sites was performed. This high throughput system, while specifically built to analyze stiffness-mediated stem cell differentiation, can be expanded to other physical cues to more broadly assess mechanical signaling and increase the pace of sensor discovery.

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Language(s): eng - English
 Dates: 2016-10-10
 Publication Status: Published in print
 Pages: 10
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Degree: -

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Title: Integrative Biology
  Abbreviation : Integr. Biol.
Source Genre: Journal
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Publ. Info: Cambridge, England : Royal Society of Chemistry
Pages: - Volume / Issue: 8 (10) Sequence Number: - Start / End Page: 1049 - 1058 Identifier: ISSN: 1757-9694
CoNE: https://pure.mpg.de/cone/journals/resource/1757-9694