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Isolation, characterization and osteogenic differentiation of adipose-derived stem cells: from small to large animal models

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Abstract

One of the most important issues in orthopaedic surgery is the loss of bone resulting from trauma, infections, tumours or congenital deficiency. In view of the hypothetical future application of mesenchymal stem cells isolated from human adipose tissue in regenerative medicine, we have analysed and characterized adipose-derived stem cells (ASCs) isolated from adipose tissue of rat, rabbit and pig. We have compared their in vitro osteogenic differentiation abilities for exploitation in the repair of critical osteochondral defects in autologous pre-clinical models. The number of pluripotent cells per millilitre of adipose tissue is variable and the yield of rabbit ASCs is lower than that in rat and pig. However, all ASCs populations show both a stable doubling time during culture and a marked clonogenic ability. After exposure to osteogenic stimuli, ASCs from rat, rabbit and pig exhibit a significant increase in the expression of osteogenic markers such as alkaline phosphatase, extracellular calcium deposition, osteocalcin and osteonectin. However, differences have been observed depending on the animal species and/or differentiation period. Rabbit and porcine ASCs have been differentiated on granules of clinical grade hydroxyapatite (HA) towards osteoblast-like cells. These cells grow and adhere to the scaffold, with no inhibitory effect of HA during osteo-differentiation. Such in vitro studies are necessary in order to select suitable pre-clinical models to validate the use of autologous ASCs, alone or in association with proper biomaterials, for the repair of critical bone defects.

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Acknowledgments

The authors are grateful to Dr. Alessandro Addis, Carlo Sala, Giuseppe Rossoni, Patrizia Rosa and Permedica S.p.a. (Merate, LC, Italy) for providing us with experimental material and to Filippo Perbellini and Giuliana Gatti for their invaluable help.

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Authors and Affiliations

  1. Department of Medical Pharmacology, Faculty of Medicine, Università degli Studi di Milano, Via Vanvitelli 32, 20129, Milan, Italy

    Elena Arrigoni, Silvia Lopa, Deborah Stanco & Anna T. Brini

  2. IRCCS Galeazzi Orthopaedic Institute, Milan, Italy

    Laura de Girolamo

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  1. Elena Arrigoni
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  2. Silvia Lopa
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  3. Laura de Girolamo
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  4. Deborah Stanco
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  5. Anna T. Brini
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Corresponding author

Correspondence to Anna T. Brini.

Additional information

This study was partially supported by grants from PRIN 2006 (area 09, prot. 2006091907_003, Italian Ministry of University and Research), FIRST 2007 and RF-IOG-2007-656853.

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Arrigoni, E., Lopa, S., de Girolamo, L. et al. Isolation, characterization and osteogenic differentiation of adipose-derived stem cells: from small to large animal models. Cell Tissue Res 338, 401–411 (2009). https://doi.org/10.1007/s00441-009-0883-x

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  • DOI: https://doi.org/10.1007/s00441-009-0883-x

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