Research ArticleCell specific differences between human adipose-derived and mesenchymal–stromal cells despite similar differentiation potentials
Introduction
Adipose tissue appears as a suitable reservoir of regenerative cells as this adult tissue is abundant and easy to sample with no ethic limitation. Recent works from independent groups state that adipose tissue hosts cells able to display various differentiation potentials in vitro and in vivo [1], [2], [3], [4]. Such cells can be isolated from the stroma vascular fraction (SVF) obtained after adipose tissue digestion that may be used either freshly prepared or after sub-culture [2], [4], [5], [6], [7]. In this last case, cultured cells represent a particular cell subpopulation, restricted to the adherent cell fraction of SVF, termed ADSC (adipose-derived stromal cells).
ADSC are often called adipose-derived mesenchymal stem cells (MSC), as MSC represent an archetype of adult stem cells. MSC are classically isolated from bone marrow (BM) aspirates and are identified as a multipotent cell population in the adult organism able to be induced to express adipogenic, osteogenic and chondrogenic markers [8], [9], [10], [11] and especially studied in the context of bone/cartilage regeneration in vivo [12], [13], [14]. Although stromal cells from BM and adipose tissue seem to be closely related, notable differences were reported [6], [15], [16], [17], [18], [19], but no systematic and extensive parallel comparison between these cells has yet been performed. According to the large perspective opened by the easy access to an abundant source of regenerative cells, such study seems requested.
The objective of this study was to realize such comparison on the biological characteristics of ADSC and MSC both in vitro and in vivo. Indeed, two dimensional gel electrophoresis and real-time PCR based Taqman Low Density Array (TLDA) covering differentiation, stemness and signalling pathways, were used to establish, respectively, a proteomic and transcriptional profile of both cell populations. In addition, we analysed their phenotype as well as their differentiation potential in vitro. In vivo, cells were injected into the intra-articular space of mouse knee joints to determine their capacity to differentiate in an in vivo cartilaginous environment.
Section snippets
Cell culture
The SVF was isolated from human adipose tissue as previously described [4]. Human subcutaneous fat was obtained from healthy patients undergoing abdominal dermolipectomy in the department of Plastic Surgery of Toulouse Rangueil Hospital (Toulouse, France). The patients were between 20 and 50 years old and their BMI never over 30. SVF cells were plated overnight in DMEM-F12 (1:1) medium supplemented with 10% FBS, 100 μg/ml pantothenic acid, 100 μM ascorbic acid, 16 μM biotin, 250 μg/ml
Phenotypic characterization of ADSC and MSC
After isolation, both MSC and ADSC exhibited fibroblast-like cell morphology. Both cells were positive for known mesenchymal markers such as CD73 and CD90 and negative for the haematopoietic marker CD45 (Fig. 1). As previously described [4], [5], [9], [23], [24]CD44 and CD105 were also expressed by both cell types, while CD31 was not detected (data not shown). MSC expressed CD106 (VCAM1), an adhesion molecule found on vascular cells that mediates the binding of haematopoietic progenitors on MSC
Acknowledgments
Caton D. and Roche S. as well as the study are financially supported by the European Community (Key action LSH 1.2.4-3, Integrated project: “Adult mesenchymal stem cells engineering for connective tissue disorders. From the bench to the bed side”, Contract no: 503161), the Cell and Gene Therapy program from Région Midi-Pyrénées (#03011999), and the Etablissement Français du Sang.
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These authors contributed equally to the work.