Fax +41 61 306 12 34 E-Mail karger@karger.ch www.karger.com Original Paper Cells Tissues Organs 2009;189:373–381 DOI: 10.1159/000154270 Ascorbic Acid Enhances Adipogenesis of Bone Marrow-Derived Mesenchymal Stromal Cells Barbara Weiser Florian Sommer Markus Neubauer Annina Seitz Joerg Tessmar Achim Goepferich Torsten Blunk Department of Pharmaceutical Technology, Institute of Pharmacy, University of Regensburg, Regensburg, Germany The presented results demonstrate AA as a potent medium component able to enhance adipogenic conversion of BMSCs, especially when administered during cell propaga- tion. Copyright © 2008 S. Karger AG, Basel Introduction One strategy in adipose tissue engineering is to gener- ate transplantable constructs seeded with adipocyte pre- cursor cells [Weiser et al., 2005; Gomillion et al., 2006]. Key Words Ascorbic acid  Adipogenesis  Mesenchymal stromal cells  Differentiation  Collagen Abstract A prerequisite to successfully engineer cell-based adipose tissue surrogates is the evaluation of in vitro culture condi- tions that facilitate expansion of primary precursor cells un- der retention of their adipogenic potential and that enable a large fraction of the heterogeneous cell pool to undergo adipogenesis upon respective stimuli. Ascorbic acid (AA) was reported to enhance differentiation of precursor cells into various mesenchymal cell types. Thus, the aim of the current study was to evaluate the influence of AA on hor- monally induced adipogenesis of bone marrow-derived mesenchymal stromal cells (BMSCs) in vitro when supple- mented during cell propagation and/or adipogenic differen- tiation. BMSCs were isolated from rat bone marrow, propa- gated, and hormonally induced to undergo adipogenesis. Supplementation of AA from the time of induction increased the fraction of BMSCs differentiating into adipocytes and glycerol-3-phosphate dehydrogenase activity up to 2-fold. Furthermore, administration of AA already during propaga- tion had an even larger effect with an up to 8-fold increase in adipogenic markers. Assessment of collagen accumula- tion suggested that the observed effects might be attribut- ed to an enhanced collagen synthesis during propagation. Accepted after revision: March 17, 2008 Published online: September 4, 2008 Dr. Torsten Blunk, Department of Pharmaceutical Technology Institute of Pharmacy, University of Regensburg Universitaetsstrasse 31, DE–93040 Regensburg (Germany) Tel. +49 941 943 4920, Fax +49 941 943 4807 E-Mail torsten.blunk@chemie.uni-regensburg.de © 2008 S. Karger AG, Basel 1422–6405/09/1896–0373$26.00/0 Accessible online at: www.karger.com/cto Abbreviations used in this paper AA ascorbic acid ADSCs adipose tissue-derived stromal cells BMSCs bone marrow-derived mesenchymal stromal cells BSA bovine serum albumin CCD charge-coupled device DHAP dihydroxyacetone phosphate DMEM Dulbecco’s Modified Eagle Medium EDTA ethylenediamine tetraacetic acid FBS fetal bovine serum GPDH glycerol 3-phosphate dehydrogenase IBMX 3-isobutyl-1-methylxanthine NADH nicotinamide adenine dinucleotide PBS phosphate-buffered saline