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Additional file 1. Supplementary Material and Method.

Additional file 2. Fig. S1: BrdU staining of Lipogems derived MSC after SF stimulation for 24 hours. SF stimulation showed approximately twofold increased fluorescence compared to untreated cells, resulting in 63,3% of positive cells when SF is added to cell culture versus 27.0% obtained in untreated cells. Fig. S2: In-Cell Western quantification of the relative intracellular level of HOXB7 (Fig S2A) and bFGF (Fig S2B) protein in Lipogems derived MSC after 1 and 5 days of stimulation with SF. At 1 day of SF stimulation, the level protein of HOXB7 decreases in a statistically significant manner. Indeed, at 5 days no significant effect on HOXB7 protein expression were observed when SF was added. Moreover, untreated cells decreased the levels of HOXB7 at 5 days of culture (p-value

Link 4.1 Overview of the main pre-clinical findings on the impact of wild-type MSC in liver diseases. Link 4.2 Overview of the main pre-clinical findings on the impact of gene-modified MSC in liver diseases. (DOCX 22 kb)

Link 1.1 Overview of the main experimental findings on the impact of wild-type MSC in diseases associated with the central and peripheral nervous system. Link 1.2 Overview of the main experimental findings on the impact of gene-modified MSC in diseases associated with the central and peripheral nervous system. (DOCX 34 kb)

Link 7.1 Overview of the main pre-clinical findings on the impact of wild-type MSC in skeletal diseases. Link 7.2 Overview of the main pre-clinical findings on the impact of gene-modified MSC in skeletal diseases. (DOCX 24 kb)

Link 6.1 Overview of the main pre-clinical findings on the impact of wild-type MSC in renal diseases. Link 6.2 Overview of the main pre-clinical findings on the impact of gene-modified MSC in renal diseases. (DOCX 29 kb)

Link 5.1 Overview of the main pre-clinical findings on the impact of wild-type MSC in pancreatic diseases. Link 5.2 Overview of the main pre-clinical findings on the impact of gene-modified MSC in pancreatic diseases. (DOCX 20 kb)

Link 2.1 Overview of the main pre-clinical findings on the impact of wild-type MSC in heart diseases. Link 2.2 Overview of the main pre-clinical findings on the impact of gene-modified MSC in heart diseases. (DOCX 39 kb)

Link 3.1 Overview of the main pre-clinical findings on the impact of wild-type MSC in lung diseases. Link 3.2 Overview of the main pre-clinical findings on the impact of gene-modified MSC in lung diseases. (DOCX 24 kb)

The resorption rate of autologous fat transfer (AFT) is 40–60% of the implanted tissue, requiring new surgical strategies for tissue reconstruction. We previously demonstrated in a rabbit model that AFT may be empowered by adipose-derived mesenchymal stromal/stem cells (AD-MSCs), which improve graft persistence by exerting proangiogenic/anti-inflammatory effects. However, their fate after implantation requires more investigation. We report a xenograft model of adipose tissue engineering in which NOD/SCID mice underwent AFT with/without human autologous AD-MSCs and were monitored for 180 days (d). The effect of AD-MSCs on AFT grafting was also monitored by evaluating the expression of CD31 and F4/80 markers. Green fluorescent protein-positive AD-MSCs (AD-MSC-GFP) were detected in fibroblastoid cells 7 days after transplantation and in mature adipocytes at 60 days, indicating both persistence and differentiation of the implanted cells. This evidence also correlated with the persistence of a higher graft weight in AFT-AD-MSC compared to AFT alone treated mice. An observation up to 180 d revealed a lower resorption rate and reduced lipidic cyst formation in the AFT-AD-MSC group, suggesting a long-term action of AD-MSCs in support of AFT performance and an anti-inflammatory/proangiogenic activity. Together, these data indicate the protective role of adipose progenitors in autologous AFT tissue resorption.

The complexity of the central nervous system (CNS) requires researchers to consider all the variables linked to the interaction between the different cell inhabitants. On this basis, any in vitro study of the physiological and pathological processes regarding the CNS should consider the balance between the standardization of the assay and the complexity of the cellular system which mimics the in vivo microenvironment. One of the main structural and functional components of the CNS is the oligodendrocyte precursor cell (OPC), responsible for developmental myelination and myelin turnover and repair during adulthood following differentiation into mature oligodendrocytes. In the present brief research report, we describe a 3D culture tool (VITVO) based on an inert and biocompatible synthetic polymer material scaffold, functionalized with laminin coating, and tested as a new culture microenvironment for neural stem/precursor cell (NSPC) differentiation compared to standard 2D cultures. NSPCs spontaneously differentiate in the three neural lineages (neurons, astrocytes and OPCs), identified by specific markers, along the fibers in the 3D structure. Analysis of the mRNA levels for lineage differentiation markers reveals a higher expression compared to those seeded on a 2D surface, suggesting an acceleration of the differentiation process. We then focused on the oligodendroglial lineage, showing that in VITVO, mature oligodendrocytes exhibit a myelinating morphology, proven by 3D image elaboration, linked to a higher expression of mature oligodendrocyte markers. This preliminary study on an innovative 3D culture system is the first robust step in producing new microenvironment-based strategies to investigate in vitro OPC and oligodendrocyte biology.

Pancreatic ductal adenocarcinoma (PDAC) is still one of the most aggressive adult cancers with an unacceptable prognosis. For this reason novel therapies accounting for PDAC peculiarities, such as the relevant stromal reaction, are urgently needed. Here adipose mesenchymal stromal/stem cells (AD-MSC) have been armed to constantly release a soluble trimeric and multimeric variant of the known anti-cancer TNF-related apoptosis-inducing ligand (sTRAIL). This cancer gene therapy strategy was in vitro challenged demonstrating that sTRAIL was thermally stable and able to induce apoptosis in the PDAC lines BxPC-3, MIA PaCa-2 and against primary PDAC cells. sTRAIL released by AD-MSC relocated into the tumor stroma was able to significantly counteract tumor growth in vivo with a significant reduction in tumor size, in cytokeratin-7+ cells and by an anti-angiogenic effect. In parallel, histology on PDAC specimens form patients (n = 19) was performed to investigate the levels of TRAIL DR4, DR5 and OPG receptors generating promising insights on the possible clinical translation of our approach. These results indicate that adipose MSC can very efficiently vehicle a novel TRAIL variant opening unexplored opportunities for PDAC treatment.

Human aging is associated with a decrease in tissue functions combined with a decline in stem cells frequency and activity followed by a loss of regenerative capacity. The molecular mechanisms behind this senescence remain largely obscure, precluding targeted approaches to counteract aging. Focusing on mesenchymal stromal/stem cells (MSC) as known adult progenitors, we identified a specific switch in miRNA expression during aging, revealing a miR-196a upregulation which was inversely correlated with MSC proliferation through HOXB7 targeting. A forced HOXB7 expression was associated with an improved cell growth, a reduction of senescence, and an improved osteogenesis linked to a dramatic increase of autocrine basic fibroblast growth factor secretion. These findings, along with the progressive decrease of HOXB7 levels observed during skeletal aging in mice, indicate HOXB7 as a master factor driving progenitors behavior lifetime, providing a better understanding of bone senescence and leading to an optimization of MSC performance. Stem Cells 2015;33:939–950