Microbioreactor array screening of Wnt modulators and microenvironmental factors in osteogenic differentiation of Mesenchymal progenitor cells

Frith, Jessica E., Titmarsh, Drew M., Padmanabhan, Harish and Cooper-White, Justin J. (2013) Microbioreactor array screening of Wnt modulators and microenvironmental factors in osteogenic differentiation of Mesenchymal progenitor cells. PLoS One, 8 12: e82931.1-e82931.15. doi:10.1371/journal.pone.0082931


Author Frith, Jessica E.
Titmarsh, Drew M.
Padmanabhan, Harish
Cooper-White, Justin J.
Title Microbioreactor array screening of Wnt modulators and microenvironmental factors in osteogenic differentiation of Mesenchymal progenitor cells
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2013-12-01
Sub-type Article (original research)
DOI 10.1371/journal.pone.0082931
Open Access Status DOI
Volume 8
Issue 12
Start page e82931.1
End page e82931.15
Total pages 15
Place of publication San Francisco, CA, United States
Publisher Public Library of Science
Language eng
Abstract Cellular microenvironmental conditions coordinate to regulate stem cell populations and their differentiation. Mesenchymal precursor cells (MPCs), which have significant potential for a wide range of therapeutic applications, can be expanded or differentiated into osteo- chondro- and adipogenic lineages. The ability to establish, screen, and control aspects of the microenvironment is paramount if we are to elucidate the complex interplay of signaling events that direct cell fate. Whilst modulation of Wnt signaling may be useful to direct osteogenesis in MPCs, there is still significant controversy over how the Wnt signaling pathway influences osteogenesis. In this study, we utilised a full-factorial microbioreactor array (MBA) to rapidly, combinatorially screen several Wnt modulatory compounds (CHIR99021, IWP-4 and IWR-1) and characterise their effects upon osteogenesis. The MBA screening system showed excellent consistency between donors and experimental runs. CHIR99021 (a Wnt agonist) had a profoundly inhibitory effect upon osteogenesis, contrary to expectations, whilst the effects of the IWP-4 and IWR-1 (Wnt antagonists) were confirmed to be inhibitory to osteogenesis, but to a lesser extent than observed for CHIR99021. Importantly, we demonstrated that these results were translatable to standard culture conditions. Using RT-qPCR of osteogenic and Wnt pathway markers, we showed that CHIR exerted its effects via inhibition of ALP and SPP1 expression, even though other osteogenic markers (RUNX2, MSX2, DLX, COL1A1) were upregulated. Lastly, this MBA platform, due to the continuous provision of medium from the first to the last of ten serially connected culture chambers, permitted new insight into the impacts of paracrine signaling on osteogenic differentiation in MPCs, with factors secreted by the MPCs in upstream chambers enhancing the differentiation of cells in downstream chambers. Insights provided by this cell-based assay system will be key to better understanding signaling mechanisms, as well as optimizing MPC growth and differentiation conditions for therapeutic applications.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
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