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Effects of GSK3 inhibitors on in vitro expansion and differentiation of human adipose-derived stem cells into adipocytes

Laure-Emmanuelle Zaragosi, Brigitte Wdziekonski, Coralie Fontaine, Phi Villageois, Pascal Peraldi and Christian Dani*

Author Affiliations

Institute of Signalling, Biology of Development and Cancer, University of Nice Sophia- Antipolis, CNRS 6543, 28 avenue Valrose, 06100 Nice, France

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BMC Cell Biology 2008, 9:11  doi:10.1186/1471-2121-9-11

Published: 13 February 2008

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Additional File 1:

Activation of β-catenin pathway by BIO. In order to evaluate the activation of the β-catenin pathway, hMADS cells were nucleofected as described in Zaragosi et al. [14] with a plasmid carrying firefly luciferase under the control of a Tcf/Lef response element (TopLuc). An inactive version of the response element was used as a control (FopLuc). Twenty four hours after nucleofection, cells were treated with BIO and 24 h after BIO treatment, cells were analyzed for luciferase expression. Renilla luciferase was co-nucleofected and used for normalization. Data indicated that β-catenin pathway was activated in hMADS cells after inhibiting GSK3.

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Additional File 2:

Morphology of hMADS3 cells after treatment with GSK3 inhibitors. Cells were maintained in medium supplemented with 0.5% FCS in the absence (Control) or presence of 0.5 μM BIO or 20 mM LiCl for 5 days.

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Additional File 3:

Impact on differentiation of GSK3 inhibition during cell proliferation. hMADS cells were maintained in the absence or presence of BIO or MeBio for 5 days. Then, cells were collected and plated at high cell density without any GSK3 inhibitor. Two days after cells reached confluence and were induced to undergo differentiation into adipocytes. GPDH activity was quantified seven days after induction of differentiation.

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Additional File 4:

Primer sequences used for quantitative PCR. Description: Primers sequences were designed using Primer Express software (Applied Biosystems, France).

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