Abstract
Identification of a suitable cell source and bioactive agents guiding cell differentiation towards tenogenic phenotype represents a prerequisite for advancement of cell-based therapies for tendon repair. Human adipose-derived stem cells (hASCs) are a promising, yet intrinsically heterogenous population with diversified differentiation capacities. In this work, we investigated antigenically-defined subsets of hASCs expressing markers related to tendon phenotype or associated with pluripotency that might be more prone to tenogenic differentiation, when compared to unsorted hASCs. Subpopulations positive for tenomodulin (TNMD+ hASCs) and stage specific early antigen 4 (SSEA-4+ hASCs), as well as unsorted ASCs were cultured up to 21 days in basic medium or media supplemented with TGF-β3 (10 ng/ml), or GDF-5 (50 ng/ml). Cell response was evaluated by analysis of expression of tendon-related markers at gene level and protein level by real time RT-PCR, western blot, and immunocytochemistry. A significant upregulation of scleraxis was observed for both subpopulations and unsorted hASCs in the presence of TGF-β3. More prominent alterations in gene expression profile in response to TGF-β3 were observed for TNMD+ hASCs. Subpopulations evidenced an increased collagen III and TNC deposition in basal medium conditions in comparison with unsorted hASCs. In the particular case of TNMD+ hASCs, GDF-5 seems to influence more the deposition of TNC. Within hASCs populations, discrete subsets could be distinguished offering varied sensitivity to specific biochemical stimulation leading to differential expression of tenogenic components suggesting that cell subsets may have distinctive roles in the complex biological responses leading to tenogenic commitment to be further explored in cell based strategies for tendon tissues.
| Original language | English |
|---|---|
| Pages (from-to) | 2204-2217 |
| Number of pages | 14 |
| Journal | Journal of Tissue Engineering and Regenerative Medicine |
| Volume | 13 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published - 1 Dec 2019 |
Funding
The authors would like to acknowledge Plastic Surgery Department of Hospital da Prelada (Porto, Portugal) for providing lipoaspirate samples. The authors acknowledge the financial support from the European Union Framework Programme for Research and Innovation HORIZON 2020, under the Tendon Therapy Train Marie Skłodowska‐Curie Innovative Training Network grant agreement No. 676338, the TEAMING Grant agreement No 739572 ‐ The Discoveries CTR, and the Achilles Twinning Project No. 810850. Authors acknowledge also the European Research Council CoG MagTendon No. 772817, the FCT (Fundação para a Ciência e a Tecnologia) Project MagTT PTDC/CTM‐CTM/29930/2017 (POCI‐01‐0145‐FEDER‐29930), and the Project NORTE‐01‐0145‐FEDER‐000021: “Accelerating tissue engineering and personalized medicine discoveries by the integration of key enabling nanotechnologies, marine‐derived biomaterials and stem cells”, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). ADE and JdB acknowledge the financial contribution of the Dutch province of Limburg in the LINK (FCL67723; “Limburg INvesteert in haar Kenniseconomie”) knowledge economy project.
Keywords
- adipose derived stromal/stem cells, subpopulation, tenogenic differentiation, tenomodulin, transforming growth factor beta 3
- stem cells
- tenomodulin
- tenogenic differentiation
- subpopulation
- transforming growth factor beta 3
- adipose derived stromal