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Culture-expanded human dermal stem cells exhibit donor to donor differences in cAMP generation

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Abstract

Stem cell techniques have facilitated a number of potential uses for such cells in cell therapy and drug development. Studies of the cAMP/protein kinase A (PKA) pathway are widely employed to investigate the effects of a large variety of substances. We assayed the cAMP pathway in human skin-derived mesenchymal stem cells (S-MSC) to evaluate donor to donor variations in response to pharmacological manipulations in vitro. Immunophenotyping of S-MSC revealed that, in general, 95% of S-MSCs were positive for CD90, CD73 and CD105 and negative for the expression of haemopoetic markers CD14, CD45 and human leukocyte antigen-DR (HLA-DR). Nevertheless, fluctuations occurred in basal cAMP levels from 5 pmol/mg to 18 pmol/mg. Total cAMP response element binding protein (CREB) concentrations ranged from 0.8 ng/ml to 1 ng/ml, whereas the proportions of phospho-CREB versus total CREB differed between the cell lines. Basic fibroblast growth factor (FGF-2) and epidermal growth factor (EGF) stimulated cAMP generation, whereas leukaemia inhibiting factor reduced some of their effects. Forskolin (0.05 and 1 mM) acted in synergy with FGF-2 and EGF; however, it caused pronounced donor to donor differences in the increase of cAMP and phospho-CREB levels. Additionally, dibutyryl-cAMP caused significant donor to donor variations in cell proliferation, possibly indicating a change of cell differentiation status. We speculate that similar donor diversity might be observed after cell stimulation with various Gs-protein-coupled receptor ligands. Heterogeneity of donor cell responses to stimulation of the cAMP pathway indicates the need for wide safety margins for S-MSC use in drug screening; nevertheless, knowledge of this heterogeneity might be useful for the design of donor-specific cell therapy.

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Authors and Affiliations

  1. Faculty of Medicine, University of Latvia, Sarlotes Street 1a, Riga, LV-1001, Latvia

    Kaspars Jekabsons, Una Riekstina, Vadims Parfejevs, Anete Laizane, Marta Pavasare, Nelda Lencberga, Baiba Jansone & Ruta Muceniece

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  1. Kaspars Jekabsons
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  2. Una Riekstina
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  3. Vadims Parfejevs
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  4. Anete Laizane
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  5. Marta Pavasare
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  6. Nelda Lencberga
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  7. Baiba Jansone
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  8. Ruta Muceniece
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Correspondence to Ruta Muceniece.

Additional information

This work was supported by European Social Foundation 2009/0217/1DP/1.1.1.2.0/09/APIA/VIAA/031 and Latvian Science Council grant no. 1037.

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Jekabsons, K., Riekstina, U., Parfejevs, V. et al. Culture-expanded human dermal stem cells exhibit donor to donor differences in cAMP generation. Cell Tissue Res 345, 253–263 (2011). https://doi.org/10.1007/s00441-011-1203-9

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  • DOI: https://doi.org/10.1007/s00441-011-1203-9

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