Abstract
Background
Unraveling the intricate and tightly regulated process of adipogenesis, involving coordinated activation of transcription factors and signaling pathways, is essential for addressing obesity and related metabolic disorders. The molecular pathways recruited by mesenchymal stem cells (MSCs) during adipogenesis are also dependent on the different sources of the cells and genetic backgrounds of donors, which contribute to the functional heterogeneity of the stem cells and consequently affect the developmental features and fate of the cells.
Methods
In this study, the alteration of transcripts during differentiation of synovial mesenchymal stem cells (SMSCs) derived from fibrous synovium (FS) and adipose synovial tissue (FP) of two pig breeds differing in growth performance (German Landrace (DL)) and fat deposition (Angeln Saddleback (AS)) was investigated. SMSCs from both tissues and breeds were stimulated to differentiate into adipocytes in vitro and sampled at four time points (day 1, day 4, day 7 and day 14) to obtain transcriptomic data.
Results
We observed numerous signaling pathways related to the cell cycle, cell division, cell migration, or cell proliferation during early stages of adipogenesis. As the differentiation process progresses, cells begin to accumulate intracellular lipid droplets and changes in gene expression patterns in particular of adipocyte-specific markers occur. PI3K-Akt signaling and metabolic pathways changed most during adipogenesis, while p53 signaling and ferroptosis were affected late in adipogenesis. When comparing MSCs from FS and FP, only a limited number of differentially expressed genes (DEGs) and enriched signaling pathways were identified. Metabolic pathways, including fat, energy or amino acid metabolism, were highly enriched in the AS breed SMSCs compared to those of the DL breed, especially at day 7 of adipogenesis, suggesting retention of the characteristic metabolic features of their original source, demonstrating donor memory in culture. In contrast, the DL SMSCs were more enriched in immune signaling pathways.
Conclusions
Our study has provided important insights into the dynamics of adipogenesis and revealed metabolic shifts in SMSCs associated with different cell sources and genetic backgrounds of donors. This emphasises the critical role of metabolic and genetic factors as important indications and criteria for donor stem cell selection.
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Background
The mesenchymal stem cells (MSCs) are multipotent progenitors cells that can differentiate into a variety of cell types, including bone cells, cartilage cells, and fat cells [1]. Adipogenesis, the differentiation of mesenchymal stem cells into adipocytes, entails activating transcription factors and signaling pathways. It comprises a two-step process: stem cell determination and preadipocyte differentiation [2,3,4]. Briefly, at the beginning of adipogenesis is the signaling of bone morphogenetic proteins (BMPs), which belong to the transforming growth factor β (TGF-β) superfamily, a family of proteins that play a role in the conversion of pluripotent stem cells into the adipocyte lineage [5]. Then it is often portrayed as a cascade of regulatory events, the first wave involving CCAAT/enhancer-binding protein β (C/EBPβ) and C/EBPδ and sterol-regulatory element binding protein 1 (SREBP1), and these transcription factors being involved in the activation of the second wave, which includes C/EBPα and PPARγ, which coordinately activate the transcription of genes that give rise to the adipocyte phenotype [4].
Pigs, due to their physiological similarities to humans, serve as valuable cellular models for advancing stem cell therapies, regenerative medicine, and transplantation [6, 7]. Porcine MSCs are utilized as large animal models in regenerative medicine, preclinical studies, and transplantation for both human and veterinary applications, benefiting the livestock industry [7,8,9]. Comprehensive understanding of molecular changes during MSC transition from self-renewal to differentiation, particularly in pig adipogenesis, and related technologies, contributes to diverse experimental research.
Various source of tissue for derived MSCs influence functional properties of the cells was reported [10,5.
Data availability
The expression datasets for this study can be found in the Gene Expression Omnibus public repository with the GEO accession number (GSE232501: GSM7349177-GSM7349248).
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Acknowledgements
We would like to express special thanks to Joana Bittner, Nicole Gentz, and Annette Jugert for their commitment and excellent technical assistance.
Funding
The Research Institute for Farm Animal Biology (FBN) provided own matched funding. The publication of this article was funded by the Open Access Fund of the Research Institute for Farm Animal Biology (FBN). Open Access funding enabled and organized by Projekt DEAL.
Open Access funding enabled and organized by Projekt DEAL.
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Performed data analysis, data curation, writing original draft, S.P.; Conceived and designed P.S., and S. P.; Conceptualization, supervision, help analysis data, S.L and S.P.; writing, review & editing, W.K., M.O., H.R., and K.W.; All authors have read and agreed to the published version of the manuscript.
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The study is reported in accordance with ARRIVE guidelines. The animals were obtained from the experimental pig unit of the Research Institute for Farm Animal Biology, Dummerstorf, Germany and were not part of an animal experiment. Animal husbandry and slaughter followed the guidelines set by the Animal Care Committee the State of Mecklenburg-Western Pomerania, Germany, based on the German Law of Animal Protection. The slaughterhouse is approved by the European Union and the German quality management system QS (MV21212). The handling and killing of the animals were carried out in accordance with applicable laws, relevant guidelines and ethical regulations, and with veterinary inspection certifying the health of the animals.
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Ponsuksili, S., Siengdee, P., Li, S. et al. Effect of metabolically divergent pig breeds and tissues on mesenchymal stem cell expression patterns during adipogenesis. BMC Genomics 25, 407 (2024). https://doi.org/10.1186/s12864-024-10308-z
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DOI: https://doi.org/10.1186/s12864-024-10308-z