Adipose-derived Stem Research Articles

EfficientIn VitroDifferentiation of Adipose Tissue-Derived Mesenchymal Stem Cells Into the Cardiomyocyte Using Plant-Derived Natural Compounds

Various degenerative diseases, traumatic injuries and cancers are almost hard to treat with conventional therapies, causing death or at least permanent disability. The use of multipotent adipose-derived mesenchymal stem cells (ADMSCs) for cell-based therapeutic applications has lately drawn elevated attention primarily due to their potential for differentiation into mesodermal lineages, which can finally be converted into cardiomyocytes to replace present invasive methods used to treat cardiac diseases. However, some reports hindered their clinical implementation due to concerns about efficacy scalability and reproducibility. In this research, we tested the impacts of using multiple natural small molecules and plant extract on the effectiveness of differentiating human ADMSCs into the definitive mesoderm lineage and cardiac progenitors. First, mesenchymal stem cells separated from human adult fat tissue, are propagated and characterized using flow cytometry and appropriate markers for this purpose. Second, passage cells are transferred to 24 well plates and treated with various plant-derived small molecules — primarily one of the most preserved intracellular signals, WNT/[Formula: see text]-catenin stimulators including resveratrol, stilbene, and multiple plant extracts — and assessed their impacts on differentiation into the definitive mesoderm and cardiac lineages. Obtained results revealed that our suggested strategy to differentiation is more viable and safer than present approaches. Taken together, data presented here showed that in vitro differentiation using plant-derived small molecules could be a potential way to improve the effectiveness of their final differentiation into definitive cardiomyocytes for in vivo applications.

Pyridoxal-5'-Phosphate Promotes Immunomodulatory Function of Adipose-Derived Mesenchymal Stem Cells through Indoleamine 2,3-Dioxygenase-1 and TLR4/NF-κB Pathway.

Adipose-derived mesenchymal stem cells (A-MSCs) are promising cellular therapies for the treatment of immune-mediated diseases. Non-gene editing technologies can improve the immune regulatory function of A-MSCs. Our preliminary experiments revealed that an active form of vitamin B6—pyridoxal-5′-phosphate (PLP)—plays an important role in regulating gene expression and cytokine secretion in A-MSCs in vivo. To further clarify the effect of PLP on receptors and cytokines related to the immune regulatory function of A-MSCs, a series of experiments were designed to verify the relationships between PLP and A-MSCs in vitro. Initially, A-MSCs were obtained, and cytokine secretion and the expression of IDO1, NF-κB, and Toll-like receptors in PLP-stimulated A-MSCs were evaluated. In addition, coculture was used to detect A-MSCs-mediated apoptosis of CD3+CD8+ T lymphocytes. These results showed that A-MSCs stimulated with PLP were highly proliferative, consistent with their pluripotent capacity. Further, the surface receptors TLR3, TLR4, IDO1, and NF-κB were upregulated, while TLR6 was downregulated. Concurrently, A-MSCs preconditioned with PLP had the greatest inhibitory effect on CD3+CD8+ T lymphocyte proliferation, indicating that PLP altered the immune regulatory function of A-MSCs through the regulation of TLRs and IDO1 expression.

The feasibility of human adipose-derived mesenchymal stem cells induced to endothelial-like cells and the safety of induced cells intraocular application in a short period

Objective To evaluate the feasibility of differentiation of human adipose mesenchymal stem cells (hADSCs) into endothelial-like cells and the safety of induced cells intraocular application. Methods HADSCs were induced to endothelial-like cells in vitro.The experimental group was added with streptomycin, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), and the control group was added with the same amount of phosphate buffered saline (PBS), the expression of von Willebrand factor (vWF), a marker of endothelial cells, was observed in the two groups.Six C57BL/6J mice were randomly divided into experimental group and control group by using the random number method, 3 mice for each group, the induced cells were injected intravitreally into C57BL/6J mice as experimental group, and the same amount of PBS buffer was injected intravitreally into C57BL/6J mice as control group.The changes of retinal cells were observed by electron microscopy.The use and care of the animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. Results After induction of hADSCs in vitro, the endothelial cell marker was expressed.VWF immunofluorescence of the experimental group showed strong green fluorescence, and the color rendering rate was 100%.The control group showed no coloration of vWF immunofluorescence, and the color positive rate was 0.After 1 month of intravitreal injection, the retinal ganglion cells and rod cells were not degraded and necrotic. Conclusions HADSCs can differentiate into endothelial-like cells in vitro, and there is no retinal toxicity in a short-term. Key words: Adipose derived mesenchymal stem cells; Endothelium-like cells; Induced differentiation; Toxicity

Intraportally delivered stem cell spheroids localize in the liver and protect hepatocytes against GalN/LPS-induced fulminant hepatic toxicity

BackgroundSystemic inflammatory response syndrome (SIRS) is common in severe fulminant hepatic failure (FHF) and has a high mortality rate (20–50%) due to irreversible cerebral edema or sepsis. Stem cell-based treatment has emerged as a promising alternative therapeutic strategy to prolong the survival of patients suffering from FHF via the inhibition of SIRS due to their immunomodulatory effects.Methods3D spheroids of adipose-derived mesenchymal stem cells (3D-ADSC) were prepared by the hanging drop method. The efficacy of the 3D-ADSC to rescue FHF was evaluated in a d-galactosamine/lipopolysaccharide (GalN/LPS)-induced mouse model of FHF via intraportal transplantation of the spheroids.ResultsIntraportally delivered 3D-ADSC better engrafted and localized into the damaged livers compared to 2D-cultured adipose-derived mesenchymal stem cells (2D-ADSC). Transplantation of 3D-ADSC rescued 50% of mice from FHF-induced lethality, whereas only 20% of mice survived when 2D-ADSC were transplanted. The improved transplantation outcomes correlated with the enhanced immunomodulatory effect of 3D-ADSC in the liver microenvironment.ConclusionThe study shows that the transplantation of optimized 3D-ADSC can efficiently ameliorate GalN/LPS-induced FHF due to improved viability, resistance to exogenous ROS, and enhanced immunomodulatory effects of 3D-ADSC.

P315In-vivo and vitro mitochondrial transfer from adipose-derived mesenchymal stem cell to ischemic cardiomyocyte

Abstract Background Although transplantation of human Adipose-derived Mesenchymal stem cell (hADSC) shows efficacy in the treatment of ischemic cardiomyopathy, its therapeutic mechanisms have not been fully elucidated. It has been already reported that mitochondria transfer to recipient cells have impact on resistance to injury and tissue regeneration, however this phenomenon has not been elucidated in the damaged heart. Therefore, we hypothesized that ADSC transfer own mitochondria to cardiomyocytes in-vivo and in-vitro under ischemic condition, resulting in the functional recovery of cardiomyocyte. Method and result Transplantation of hADSC (group A) to the heart surface or sham operation (group C) was performed in rats that were subjected to LAD ligation 2 weeks prior to the treatment (n=10 each). The number of transplant cell was 1x106/body. Three days after transplantation, transferred hADSCs' mitochondria were observed in recipient cardiomyocytes histologically (Figure). Quantitative PCR analysis revealed that mitochondrial genome of recipient myocytes increased over time. The cardiac function assessed with echocardiography was significantly better in group A. Furthermore, live-imaging of hADSC transplantation revealed the suspected transfer of mitochondria to beating heart. In-vitro, the co-culture of rat cardiomyocytes (rCM) and hADSC was observed with time-lapse photography and demonstrated mitochondrial transfer under the hypoxic condition. The measuring the oxygen consumption rate (OCR) of these cells showed that OCR of rCM was reinforced by co-culture with hADSC conspicuously. Figure 1 Conclusion Mitochondrial transfer from hADSC to rCM was suggested in-vivo and in-vitro ischemic condition and suspected to be related to functional recovery of ischemic cardiomyocyte.

Impaired immunomodulatory ability of type 2 diabetic adipose-derived mesenchymal stem cells in regulation of inflammatory condition in mixed leukocyte reaction.

The immunomodulatory properties of type 2 diabetic patients' adipose-derived mesenchymal stem cells (D-ASCs) has not been extensively studied. In this study, we compared the immunomodulatory properties of D-ASCs and non-diabetic subjects mesenchymal stem cells (ND-ASCs) in co-culture with mixed leukocyte reaction (MLR). ASCs were isolated from adipose tissue samples of type 2 diabetic and non-diabetic subjects (age: 40-55). D-ASCs and ND-ASCs were co-cultured with two-way MLR. Peripheral blood mononuclear cells (PBMCs) proliferation ratio, protein levels of IFN-γ and IL-10, mRNA expression of COX-2, TNF-α, TGF-β1 and IL-6 genes in MLR, D-ASCs and ND-ASCs co-cultures were assessed using XTT, ELISA and Real-time qRT-PCR, respectively. PBMCs proliferation on days 2 and 4 of D-ASCs co-culture was higher than ND-ASCs co-culture of the same days (p < 0.001). IFN-γ level decreased on day 4 compared to day 2 of ND-ASCs co-culture, but its level had not changed in D-ASCs co-culture. COX-2 expression on days 2 and 4 of D-ASCs co-culture was lower than ND-ASCs co-culture of the same days (p < 0.05). The expression of TNF-α and IL-6 on days 2 and 4 of D-ASCs co-culture were higher than ND-ASCs co-culture of the same days (p < 0.001). TGF-β1 on day 4 of ND-ASCs co-culture showed a slightly higher expression than D-ASCs co-culture of the same day. Lower suppression of PBMCs proliferation, declined expression of anti-inflammatory and upregulated expression of pro-inflammatory factors in D-ASCs co-culture, indicated an impairment of these cells in modulation of the inflammatory condition.

T-Box20 inhibits osteogenic differentiation in adipose-derived human mesenchymal stem cells: the role of T-Box20 on osteogenesis

BackgroundSkeletal development and its cellular function are regulated by various transcription factors. The T-box (Tbx) family of transcription factors have critical roles in cellular differentiation as well as heart and limbs organogenesis. These factors possess activator and/or repressor domains to modify the expression of target genes. Despite the obvious effects of Tbx20 on heart development, its impact on bone development is still unknown.MethodsTo investigate the consequence by forced Tbx20 expression in the osteogenic differentiation of human mesenchymal stem cells derived from adipose tissue (Ad-MSCs), these cells were transduced with a bicistronic lentiviral vector encoding Tbx20 and an enhanced green fluorescent protein.ResultsTbx20 gene delivery system suppressed the osteogenic differentiation of Ad-MSCs, as indicated by reduction in alkaline phosphatase activity and Alizarin Red S staining. Consistently, reverse transcription-polymerase chain reaction analyses showed that Tbx20 gain-of-function reduced the expression levels of osteoblast marker genes in osteo-inductive Ad-MSCs cultures. Accordingly, Tbx20 negatively affected osteogenesis through modulating expression of key factors involved in this process.ConclusionThe present study suggests that Tbx20 could inhibit osteogenic differentiation in adipose-derived human mesenchymal stem cells.

Individual Patient Expanded Access IND of Hope Biosciences First Blood Relative Allogeneic Adipose-derived Mesenchymal Stem Cells (HB-adMSCs) for Pancreatic Cancer

Individual Patient Expanded Access IND of Hope Biosciences First Blood Relative Allogeneic Adipose-derived Mesenchymal Stem Cells (HB-adMSCs) for Pancreatic Cancer

The Hypoxic Preconditioning Effect On Senescence Cell Process In Cultured Adipose-Derived Mesenchymal Stem Cells (AMSCs)

Introduction : Stem cell therapy for myocardial regeneration is expected to increase cardiomyocyte proliferation and trigger neovascularization to improve cardiomyocytes. Mesenchymal Stem Cells (MSCs) are ideal candidates for regenerative medicine and immunotherapy. But low viability of MSCs is a major challenge in this alternative therapy. Therefore, a cytoprotective strategy is needed, one of them is hypoxic preconditioning which can significantly increase survival stem cells after being transplanted. MSCs are known to have a limited life span, after experiencing several splits MSC will enter the senescence process. It is known that hypoxia can also increase cell proliferation and differentiation potential in vitro and in vivo through the role of Octamer-4 (Oct-4) as a regulator of the pluripotency gene.&#x0D; Methods : Experimental laboratory studies (in vitro studies) using human-AMSCs which were given hypoxic preconditioning, observed as a immunocytochemistry.&#x0D; Results : The results showed that hypoxic precondition (1% O2) inhibited the senescence process. It can be seen in the lower expression of senescence in hypoxic conditions at P6, P7, P8, P9, P10 compared to normoxic ((p=0,004, p=0,001, p=0,009, p=0,013, p=0,024. There is a significant difference in the senescence expression of each passage in hypoxic and normoxic conditions with the highest expression at P10. In addition, we also observed AMSCs differentiation through the Oct-4 expression. It is showed that Oct-4 expression were higher in hypoxia compared to normoxia on P7, P8, P9, P10 (p=0,009, p=0,009, p=0,030, p=0,0001).&#x0D; Conclusions : Hypoxic preconditioning have the effect of inhibiting the senescence process on Adipose-derived MSCs (AMSCs) or prolonging their life span. The longer life span of AMSCs is also seen by higher cell differentiation potential from increased expression of Oct-4. However, the mechanism of inhibiting the senescence process in hypoxia in stem cells is still remain unknown.&#x0D; Keywords: human-Adipose derived Mesenchymal Stem Cell Cultures (h-AMSCs), Hypoxic Preconditioning, Senescence cell, Oct-4.

Adipose-Derived Mesenchymal Stem Cells as the Panacea for Degenerative Joint Diseases: Fact or Fiction?

The continuously evolving society of the third millennium requires more and more athletic skills for citizens of all countries.

Small Molecule-Based Strategy Promotes Nucleus Pulposus Specific Differentiation of Adipose-Derived Mesenchymal Stem Cells.

Adipose tissue-derived mesenchymal stem cells (ADSCs) are promising for regenerating degenerated intervertebral discs (IVDs), but the low efficiency of nucleus pulposus (NP)-specific differentiation limits their clinical applications. The Sonic hedgehog (Shh) signaling pathway is important in NP-specific differentiation of ADSCs, and Smoothened Agonist (SAG) is a highly specific and effective agonist of Shh signaling. In this study, we proposed a new differentiation strategy with the use of the small molecule SAG. The NP-specific differentiation and extracellular matrix (ECM) synthesis of ADSCs were measured in vitro, and the regenerative effects of SAG pretreated ADSCs in degenerated IVDs were verified in vivo. The results showed that the combination of SAG and transforming growth factor-β3 (TGF-β3) is able to increase the ECM synthesis of ADSCs. In addition, the gene and protein expression levels of NP-specific markers were increased by treatment with SAG and TGF-β3. Furthermore, SAG pretreated ADSCs can also improve the disc height, water content, ECM content, and structure of degenerated IVDs in vivo. Our new differentiation scheme has high efficiency in inducing NP-specific differentiation of ADSCs and is promising for stem cell-based treatment of degenerated IVDs.

P9. Dimethyloxaloylglycine (DMOG) preconditioning of adipose-derived mesenchymal stem cells as strategy for enhancing posterolateral lumbar fusion in a rat model

BACKGROUND CONTEXT Transplantation of adipose-derived mesenchymal stem cells (ADSCs) has become a promising approach for bone regeneration therapies, particularly in elderly patients. However, poor cell survival in vivo after implantation due to the ischemic tissue environment has been one of the major limitations of the effectiveness of this strategy. Hypoxia Inducible Factor-1α (HIF-1α) plays an important role in angiogenic-osteogenic coupling during bone regeneration, as it is a critical mediator of the adaptive cell response to hypoxia. DMOG treatment has been demonstrated to stabilize cellular expression of HIF-1α in cells, as well as enhance osteogenic and angiogenic capacity of MSCs. By therapeutically targeting both osteogenesis and vascularization via DMOG preconditioned MSCs, this tissue-engineering approach sought to yield superior bone quality and integration, thereby increasing rates of spinal fusion. PURPOSE We aimed to demonstrate that preconditioning adipose (ADSC)-derived (AD) MSCs with DMOG prior to implantation will prolong survival in vivo and improve spinal fusion outcomes in a rat model. STUDY DESIGN/SETTING In vivo study via a rat posterolateral spinal fusion model. PATIENT SAMPLE Syngeneic 6-8 week old Lewis rats. OUTCOME MEASURES Manual palpation scoring was performed by blinded researchers as follows: 0 = non-fused; 1 = partial fusion, some motion across operative joint; and, 2 = fused, no motion across the operated joint. MicroCT images were used to assess fusion mass volume (mm^3) via ImageJ software and determine CT fusion score (0 = non-fused; 1 = unilateral fusion; 2 = bilateral fusion). METHODS ADSCs were isolated from the inguinal fat pads of syngeneic 6-8 week old Lewis rats and cultured in vitro. When passage 1 (P1) ADSCs reached approximately 80% confluency they were pre-conditioned for 24 hours with 1 ng of DMOG (Fisher Scientific). After pre-conditioning, 2 × 10^6 ADSCs were seeded onto Vitoss (Stryker) bone graft substitute scaffolds for subsequent transplantation. Posterolateral spinal fusion surgery at L4-L5 was performed on 18 female Lewis rats divided into 2 experimental groups: [1] Vitoss+ADSCs pre-conditioned with DMOG; and, [2] Vitoss+non pre-conditioned ADSCs. Fusion was evaluated eight weeks post-surgery. Manual palpation scoring evaluated the motion across the operated joint, and MicroCT images were assessed for fusion mass volume and CT fusion. RESULTS Preliminary microCT imaging data suggest that DMOG pre-conditioned ADSCs (n=12) yielded significantly higher fusion mass volumes than non-preconditioned ADSCs (n=6) (23.49 mm^3 vs. 15.39 mm^3, respectively, p=.001). DMOG pre-conditioned ADSCs showed a trend towards higher rates of fusion and manual palpation scores compared to non-conditioned ADSCs (1.16 vs. 0.50, respectively, p=.06; and 1.25 vs. 0.66, respectively, p>.05). CONCLUSIONS In our rat posterolateral fusion model, DMOG pre-conditioned ADSCs displayed an increased fusion mass volume, CT fusion rates, and manual palpation score compared to non-conditioned ADSCs. Ongoing histological studies will evaluate whether there are any differences in the quality of bone formed within the fusion masses. Future studies will also compare whether these results are also similar in bone marrow-derived MSCs. FDA DEVICE/DRUG STATUS Dimethyloxaloylglycine (Investigational/Not approved)

Individual Patient Expanded Access IND of Hope Biosciences Autologous Adipose-derived Mesenchymal Stem Cells for Chronic Spinal Cord Injury

Individual Patient Expanded Access IND of Hope Biosciences Autologous Adipose-derived Mesenchymal Stem Cells for Chronic Spinal Cord Injury

Individual Patient Expanded Access IND of Hope Biosciences Autologous Adipose-derived Mesenchymal Stem Cells for Parkinson's Disease

Individual Patient Expanded Access IND of Hope Biosciences Autologous Adipose-derived Mesenchymal Stem Cells for Parkinson's Disease

Adipose stem cell‐derived exosomes in combination with hyaluronic acid accelerate wound healing through enhancing re‐epithelialization and vascularization

Adipose stem cell‐derived exosomes in combination with hyaluronic acid accelerate wound healing through enhancing re‐epithelialization and vascularization

Differentiation and Anti-inflammatory Potentials of Eucomis autumnalis and Pterocarpus angolensis Extracts Scaffolds in Porcine Adipose–Derived Mesenchymal Stem Cells

The key ingredients for tissue engineering and regenerative medicine are type of cells, growth factors, and biological engineering materials/scaffolds. Primary porcine adipose–derived mesenchymal stem cells (pADMSCs) were characterized using fluorescence-activated cell sorting analysis for mesenchymal surface markers of CD44, CD90, and CD105 (≥ 85%). Cell attachment was evaluated using scanning electron microscope (SEM) and immunofluorescence. MTT and histology assays were employed to evaluate pADMSCs viability and multi-potency differentiation respectively on Eucomis autumnalis and Pterocarpus angolensis crude extracts scaffolds. IL-6 ELISA and RT-PCR were used to access anti-inflammatory and bone/cartilage marker expressions respectively. Results showed that multi-potency differentiation of pADMSCs was high and dose dependent on groups treated with E. autumnalis and P. angolensis. SEM and immunofluorescence results revealed more cells attached after 7, 14, and 21 days on treated groups than untreated groups. Cell viability was high with significant difference of P < 0.01 in plant extract groups after 7, 14, and 21 days. The E. autumnalis crude extract significantly (P < 0.01) suppressed the protein expression of IL-6 in comparison to extracts prepared from P. angolensis and untreated groups. Collagen type II and osteopontin were significantly expressed in P. angolensis group compared to E. autumnalis and other groups. Alkaline phosphatase and osteocalcin genes were significantly expressed on days 14 and 21 in E. autumnalis compared to other groups. This study provides some evidence to support the anti-inflammatory nature of E. autumnalis and P. angolensis extract as well as proof that these plants are able to induce multi-potent differentiation of ADMSCs. Eucomis autumnalis and Pterocarpus angolensis are among the plants used by numerous traditional healers to reduce osteoarthritis symptoms and improve bone fracture healings. This work explored the potentials of plant extracts incorporated scaffolds to induce adipogenic, osteogenic, chondrogenic differentiation, and anti-inflammatory effects in porcine adipose-derived mesenchymal stem cells (pADMSCs). Our data showed that plant extract scaffolds suppressed the expression of interlukin-6 especially in E. autumnalis in our ELISA assay analysis which confirmed the anti-inflammatory nature of the plant extract scaffolds. The plant extract scaffolds also depicted osteogenic and chondrogenic support in the multi-potency differentiation of the pADMSCs. Pterocarpus angolensis extract scaffold showed high upregulated expression of collagen type II on day 21 at the highest dose of 5 mg/ml. In conclusion, plant extracts in scaffolds may provide safe, cost-effective alternative treatment for bone healing and cartilage regeneration and stimulation of bone formation by virtue of their anti-inflammatory properties. Our research work is relevant to communities that rely solely on traditional medicine for their well-being, orthopedics, pharmaceutical companies, regenerative, and translational medicine fields.

Regulation of somatostatin expression by vitamin D3 and valproic acid in human adipose-derived mesenchymal stem cells

BackgroundAdipose-derived mesenchymal stem cells (ADMSC) are non-haematopoietic, fibroblast-like multipotent progenitor cells. They have the potential for trilineage (adipocyte, chondrocyte and osteocyte) differentiation as well as differentiation into endocrine pancreatic progenitors. In diabetic or cancer therapy, somatostatin (SST) expression plays a vital role. Small molecules such as valproic acid (VPA) and micronutrients like vitamin D3 have differentiation potential in ADMSC. Therefore, the aim of this study was to investigate the role of vitamin D3 machinery and its metabolic enzymes in ADMSC. Furthermore, the reprogramming effect of vitamin D3 and VPA was evaluated on somatostatin expression in pancreatic lineage differentiation.MethodsADMSC were characterised based on their cell surface marker profile using flow cytometry. Specific adipogenic and osteogenic differentiation protocols were used in this study. Gene expression of several pluripotent, endodermal, pancreatic progenitor and pancreatic endocrine lineage markers were investigated in native ADMSC and after stimulation with different concentration of vitamin D3 for five consecutive days (0, 50, 100, 150 nM) and VPA (0.5, 1, 1.5, 2 mM) by real-time PCR. Furthermore, somatostatin expression was confirmed with ELISA and immunocytochemistry.ResultsIn ADMSC, the expression of somatostatin mRNA, the vitamin D receptor (VDR) and its metabolising enzymes 1 α-Hydroxylase, 24-Hydroxylase and 25-Hydroxylase were detected. Upon stimulation with vitamin D3, nuclear translocation of vitamin D receptor (VDR) was observed. Interestingly, the presence of vitamin D3 reduced the transcription of the somatostatin gene. By contrast, VPA treatment of cultivated ADMSC showed enhancing effect on somatostatin gene expression. No other pluripotent, endodermal, pancreatic progenitor or pancreatic endocrine lineage mRNA expression was modulated under the influence of vitamin D3 and VPA.ConclusionHuman ADMSC carry the VDR. The vitamin D metabolising enzyme 25-Hydroxylase responded to the addition of vitamin D3. Moreover, our results demonstrate that somatostatin expression in ADMSC is constitutive, partially secreted and regulated by vitamin D3 and VPA.

Immunoregulatory, proliferative and anti-oxidant effects of nanocurcuminoids on adipose-derived mesenchymal stem cells.

Curcuminoids are dietary complexes extracted from the seeds of Curcuma longa L. that contain curcumin, bisdemethoxycurcumin and desmethoxycurcumin. Curcuminoids are popular for their pleiotropic therapeutic functions, such as their anti-inflammatory and anti-oxidant effects. Nonetheless, their clinical use is associated with poor systemic bioavailability and insolubility. The nano-formulation of curcuminoids eliminates these shortcomings. In the present study, we explored immunoregulatory, proliferative and anti-oxidant effects of nanocurcuminoids on adipose-derived mesenchymal stem cells (AT-MSCs). Flow cytometry analysis and MTT assay were employed to explore the effects of nanocurcuminoids on the apoptosis and proliferation of adipose-derived MSCs (AT-MSCs). The anti-oxidant effect of nanocurcuminoids on AT-MSCs also was examined. The immune regulatory effect of nanocurcuminoids was evaluated by the flow cytometric measurement of the T regulatory (Treg) population. The expression of inflammatory and anti-inflammatory cytokines was quantified using real-time PCR. Our findings demonstrate that low concentrations of nanocurcuminoids are beneficial for MSC proliferation, protection of MSCs from apoptosis, reducing inflammatory cytokines and SOD activity. A high concentration of nanocurcuminoids increases the population of Tregs and elevates the expression of TGFβ and FOXP3 genes. The beneficial effects of nanocurcuminoids on AT-MSCs were mainly observed at low doses of nanocurcuminoids.

Epidural adipose tissue-derived mesenchymal stem cell activation induced by lung cancer cells promotes malignancy and EMT of lung cancer

BackgroundSpinal metastasis is a major challenge in patients with advanced lung cancer, but the mechanisms in the organotropism of metastasis are still unclear. Adipose-derived mesenchymal stem cells (ADSCs) exhibit cancer-promoting properties that influence the tumour microenvironment; however, there is no research on ADSCs from epidural fat thus far.MethodsIn this study, we isolated and identified ADSCs from epidural adipose tissue for the first time. We examined the activation of epidural ADSCs treated with lung cancer cell-conditioned medium by immunohistochemistry, western blot and qRT-PCR assays. The expression of interleukin (IL)-6 family cytokines in the supernatants of ADSCs were evaluated by enzyme-linked immunosorbent assay. The effects of epidural ADSCs on the growth and invasion of lung cancer cells were evaluated with the CCK-8 and Transwell assays. The expression of signal transducer and activator of transcription 3 (STAT3), matrix metalloprotease and epithelial-mesenchymal transition markers were measured by western blot assays.ResultsOur results showed that ADSCs treated with lung cancer cell-conditioned medium expressed higher levels of the myofibroblast marker α-smooth muscle actin and fibroblast activation protein than ADSCs cultured alone. Then, we found that lung cancer cells induced ADSCs to secrete high levels of IL-6 family cytokines and activate the STAT3 signalling pathway. Moreover, activated epidural ADSCs exhibited the ability to promote lung cancer cell proliferation and invasion by elevating matrix metalloprotease expression and epithelial-mesenchymal transition in cancer cells. Furthermore, blocking IL-6 can counteract the differentiation and tumour-promoting effects of ADSCs.ConclusionOur results suggest that ADSCs respond to lung cancer cells and are involved in the crosstalk between primary tumours and pre-metastatic niches in epidural fat.

Mesenchymal stem cells for the treatment of complex perianal fistulas in patients with Crohn disease.

Treatment of perianal fistulizing Crohn's disease is demanding and burdened with a high percentage of failures, which forces clinicians to search for new, more effective therapeutic options. One of these options is the use of adipose-derived mesenchymal stem cells in local administration. Due to their multipotentiality and complex mechanism of action, stem cells are the promising new therapeutic approach for the treatment-refractory complex perianal fistulas - demonstrating both high efficacy and a favorable safety profile. The paper presents current knowledge on the mechanisms of action and manner of administration of mesenchymal stem cells, as well as the effectiveness and safety of their use in the treatment of perianal Crohn's disease based on available literature.