Vitamin E and hCG enhance the immunomodulatory properties of LPS-induced mesenchymal stem/stromal cells

Mesenchymal stem/stromal cells (MSCs) are multipotent progenitors that are derived from most adult tissue as well as cord blood and placenta. MSCs are defined by their adherent nature, ability to propagate in culture and capacity to differentiate into bone fat and cartilage. However, many studies have shown that MSCs, derived from different tissues, differ both in their in situ and in vitro phenotypes. Despite abundance of MSCs studies, little is known about the molecular events that control their tissue specific nature. Two recent studies comparing MSCs derived from different tissues have now found clues to the molecular mechanisms that control the tissue specific nature ofthese cells. In the first, the superior genomic stability of adipose derived MSCs (ASCs), compared to bone marrow (BM) MSCs, was explained by reduced H19, a long non-coding RNA expression and increased p53 activity of ASCs. In the second, a compression of abdominal and subcutaneous ASCs reveals poor propagation, differentiation and migration capacities of abdominal ASCs that is explained by their increased tendency to over-accumulate reactive oxygen species (ROS) in culture. ROS over production in abdominal ASCs was shown to be controlled by the NADPH oxidase NOX1. The unique features of MSCs derived from different tissues suggest a tissue specific molecular signature arising from the tissue of origin that is retained during culture. The implications of this phenomenon on our understanding of the role and function of MSCs in situ as well as on their clinical utilization, is discussed.

Cumulative Evidence That Mesenchymal Stem Cells Promote Healing of Perianal Fistulas of Patients With Crohn's Disease–Going From Bench to Bedside

The immunomodulatory properties of mesenchymal stem/stromal cells (MSCs) make them an attractive therapeutic tool to treat chronic inflammatory diseases, such as rheumatoid arthritis or Crohn's disease. These indications are characterized by a chronic activation of immune cells that perpetuates the disease. In vitro, when adipose mesenchymal stem cells (ASCs) are cultured with T lymphocytes at the time of stimulation, their proliferation is inhibited. However, these experimental settings do not necessarily fit with what ASCs will face in inflammatory conditions in vivo, where ASCs will likely encounter and interact with already activated immune cells which might affect their immunomodulatory capacity. In most in vitro studies, MSCs have been cultured with peripheral blood mononuclear cells at the time of lymphocyte stimulation and information about the interaction between MSCs and prestimulated lymphocytes in vitro is scarce. Therefore, a better understanding of the capacity of MSCs to modulate the responses of preactivated immune cells is needed. In this study we focused on the effects of ASCs on prestimulated lymphocytes and systematically investigated the potential mechanisms involved. We report that prestimulation of T lymphocytes 48 h before the coculture with ASCs impairs the capacity of ASCs to inhibit proliferation. Preactivation of ASCs with interferon γ or the toll-like receptor ligand Poly I:C, but not other stimuli tested, enhanced the ability to inhibit the proliferation of 48 h-stimulated T lymphocytes. The inhibitory effect of ASCs was shown to be time dependent and mediated through the actual magnitude of tryptophan degradation by indoleamine 2,3-dioxygenase.

Background Corneal chemical burn is one of blinding eye diseases.Previous therapies for corneal chemical burn is limited to certain extent.However, transplantation of adipose-derived mesenchymal stem cells (ADSCs) for corneal diseases is drawing more and more attention. Objective This study was to observe the effect of rabbit ADSCs transplantation for ocular alkali burns and explore its mechanism. Methods Rabbit corneal stromal cells (CSCs) were isolated and cultured by suspended matrix method, and rabbit ADSCs were obtained and digested from inguinal fat tissue with enzyme digestion method (0.25% trypsin) and identified by flow cytometry.CSCs cocultured with ADSCs, and CSCs-induced ADSCs were identified by double-label of with immunofluorescence and reverse transcription PCR (RT-PCR). Then induced or uninduced ADSCs were inoculated on amniotic membrane to prepared ADSCs-amnion patch.Corneal alkali burn models were established in the right eyes of 60 New Zealand rabbits by placing a filter paper with 1% NaOH solution at the central cornea for 50 seconds.The models were randomized into the induced ADSCs+ amnion implanted group, the uninduced ADSCs+ amnion implanting group, amnion implanted group and model group.Corneal opacification and neovascular area were examined and corneal inflammation was graded by slit lamp microscope 1 week, 2 weeks and 1 month after surgery.The contents of CD45, interferon-γ (IFN-γ) and interleukin-10 (IL-10) in corneal homogenate as well as vascular endothelial growth factor (VEGF) in aqueous humor were detected by ELISA assay.The use and care of experiment animals followed the Statement of ARVO. Results ADSCs showed the positive responses for CD105, CD29, CD44 with the positive rate 90.23%, 88.56% and 98.88%, respectively.CSCs was positively reactive for vimentin.The double-label staining was positive after coculture of CSCs with ADSCs.Hematoxylin-eosin stain exhibited that ADSCs grew well on the amnion.Corneal porcelain opacity and a lot of new blood vessels were seen in the model group, and corneal was clear in the induced ADSCs+ amnion implanted group 1 month after surgery.The inflammatory scores were 1.65±0.18, 2.05±0.17, 2.68±0.25, 2.90±0.18, and the areas of neovasculization were (10.59±1.78), (22.58±1.63), (37.98±1.90), (45.37±1.65)mm2 respectively in the induced ADSCs+ amnion implanted group, uninduced ADSCs+ amnion implanted group, amnion implanted group and the model group.The inflammatory scores of 1 week, 2 weeks, 1 month after operation among the four groups had statistically significant differences（F=280.826, 330.172, 465.707, all at P=0.000), and the areas of neovasculization of 1 week, 2 weeks, 1 month after operation among the four groups had statistically significant differences（F=60.020, 670.811, 1 510.231, all at P=0.000), the inflammatory scores in the induced ADSCs+ amnion implanted group were remarkably lower than those of the other groups, the areas of neovasculization in the induced ADSCs+ amnion inplanted group were smaller than those of the other groups (all at P<0.01). In 1 month after surgery, the contents of CD45, IL-10, IFN-γ in cornea and VEGF in aqueous humor were statistically different among the groups（F=916.545, 1 739.358, 462.134, 129.126, all at P=0.000). Compared with the uninduced ADSCs+ amnion implanted group, amnion implanted group and the model group, CD45 and IFN-γ contents were declined, and IL-10 content was elavated in the induced ADSCs+ amnion implanted group (all at P<0.01). In addition, VEGF contents in aqueous humor were significantly lower than those in the other groups (all at P<0.01). Conclusions Rabbit CSCs-induced ADSCs amnion patch transplantation is effective for the reconstruction of ocular surface after alkali damage probably by differentiation of ADSCs into epithelial-like cell after CSCs induced.Moreover, amnion can alleviate immuno-inflammatory response and suppress neovascularization. Key words: Adipose-derived mesenchymal stem cells; Transplantation; Amniotic membrane; Alkali burn, ocular; Differentiation; Corneal stromal cells

Objective: To observe the effect of basic fibroblast growth factor (bFGF) treatment on efficacy of adipose-derived mesenchymal stem cells (ADSCs) in cirrhotic rats. Methods: A rat model of liver cirrhosis was established via intraperitoneal injection of carbon tetrachloride (CCl(4)) for 10 weeks. Thirty SD rats were randomly divided into 3 groups (n = 10): control group served as group A, and 0.5ml of phosphate-buffered saline (PBS) was transfused into the tail vein; ADSCs single-dose transplantation group served as group B, and 1×10(6) ADSCs were transplanted into the tail vein; bFGF-treated ADSCs treatment group served as group C, and 1×10(6) bFGF-treated ADSCs were transplanted through tail vein. Liver function, pathological and cytokine changes, and the in vivo survival transformation condition of the transplanted cells were measured at one week after transplantation. F test and an independent sample t test were used. Results: bFGF treatment had significantly promoted the proliferation, differentiation and overexpression of hepatocyte growth factor (HGF) in ADSCs [ADSCs single: (2 137.16 ± 261.52) pg/ml vs. ADSCs (bFGF): (4 776.23 ± 532.44) pg/ml, P < 0.05]. The bFGF-treated ADSCs treatment group had statistically significant differences in promoting the recovery of liver function [alanine aminotransferase (ALT): ADSCs single (190.8 ± 34.98) vs. ADSCs (bFGF): (117.8 ± 35.81) pg/ml; aspartate aminotransferase (AST): ADSCs single (295.2 ± 33.71) U/L vs. ADSCs (bFGF): (183.8 ± 41.29) U/L, P ​​< 0.05). There was no statistically significant difference in serum albumin levels between the control group, ADSCs single group, and ADSCs (bFGF) group. Compared with the control group, the serum albumin level of ADSCs (bFGF) group was increased significantly (P < 0.05), and the difference between the control group and the ADSCs single transplantation group in improving liver regeneration and reducing liver damage was statistically significant (P < 0.05). Masson trichrome staining showed that the percentage of the liver fibrosis area in the bFGF-treated ADSCs treatment group was 6.78% ± 0.56%, which was significantly higher than that of the control and ADSCs single dose transplantation group (7.96% ± 0.64%) (P < 0.05). Western blot analysis showed that the expression of HGF protein in the bFGF-treated ADSCs treatment group was significantly up-regulated, while the expression of α-smooth muscle actin was significantly down-regulated, and the difference was significant in the ADSCs single transplantation group. A double fluorescent staining method showed that the numbers of stem cells implanted in the liver tissue of the bFGF-treated ADSCs treatment group were higher than that of the ADSCs single transplantation group. In-vitro cell experiments confirmed that bFGF had significantly promoted the overexpression of HGF in ADSCs. Conclusion: bFGF-treated ADSCs transplantation can significantly improve liver function and fibrosis as compared with ADSCs single-dose transplantation in cirrhotic rats.

Background and objectives Adipose-derived mesenchymal stem cells (ASCs) have immunomodulatory properties, but their activity is dependent on signals provided by local microenvironment. ASCs administration in rheumatoid arthritis (RA) did not bring consistent results, while in osteoarthritis (OA) it has beneficial effects. It is likely that proinflammatory milieu of rheumatoid joint affects ASCs activity. To test this hypothesis, the function of rheumatoid ASCs (RA-ASCs) and osteoarthritic ASCs (OA-ASCs) isolated from infrapatellar fat pad of the knee joint have been analysed. Materials and methods Infrapatellar fat pads were obtained from 29 RA and 12 OA patients undergoing total knee joint replacement surgery. Then, ASCs were isolated accordingly to routinely applied procedure. ASCs were treated or not with IFNγ or TNF. The expression of indoleamine 2,3-dioxygenase and heme oxygenase 1 mRNA was measured using quantitative PCR, while ASCs secretory potential was assessed by specific ELISAs. Rheumatoid synovial fibroblasts (RA-FLS) or peripheral blood mononuclear cells (PBMCs) from healthy donors were cultured in ASC-conditioned media or co-cultured with ASCs. Next, cytokines secretion and cells proliferation were measured using ELISAs and bromodeoxyuridine/ 3 H-thymidine proliferation assays, respectively. Results RA- and OA-ASCs activity in vitro is comparable, nonetheless some differences in spontaneous secretory activity and heme oxygenase 1 mRNA expression have been observed. Unstimulated RA- and OA-ASCs slightly inhibited PBMCs proliferation and induced IL-10 secretion but increased IL-17A production and failed to suppress release of other proinflammatory factors (TNF, IFNγ, RANTES) by PBMCs. Under TNF stimulation, RA- and OA-ASCs up-regulated IL-6 and MMP-3 secretion in RA-FLS cultures and IL-17A release by PBMCs. ASCs treatment with IFNγ or TNF did not intensify their anti-inflammatory activity. Conclusions Our study demonstrates that immunosuppressive function of RA- and OA-ASCs derived from infrapatellar fat pad is impaired. Possibly, it is due to the localisation of ASCs in the site of inflammation. Moreover, TNF triggers proinflammatory properties of ASCs which indicates that when administered to the rheumatoid joint, ASCs may shift from an anti-inflammatory to proinflammatory phenotype. Acknowledgements This work was supported by the Polish National Science Centre (grant no. 2011/01/N/NZ5/00932) and by the European Union funds (European Social Fund, Human Capital Programme).

Mesenchymal stem/stromal cells preconditioning with Glibenclamide augment their immunoregulatory characteristics

<b>Objective:</b> To investigate the effect and related mechanism of apolipoprotein A5 (ApoA5) on adipogenic differentiation of human adipose-derived mesenchymal stem cells (AMSC). <b>Methods:</b> Subcutaneous adipose tissue was obtained from 40 patients undergoing abdominal surgery at our hospital from February to July 2015. After induction of human AMSC by collagenase digestion, the adipose tissue was induced to differentiate into mature adipocytes and treated with ApoA5 at 600 and 1 200 ng/ml, respectively (ApoA5 intervention groups). Cells treated without ApoA5 protein were used as control group. The cells were harvested on the 7th and 14th day of differentiation, and the following assays were performed: (1) the effect of ApoA5 on TG content was measured by a TG assay kit; (2) RT-qPCR assay was used to detect the effect of ApoA5 on aP2 and FAS mRNA expression; (3) the effect of ApoA5 on the expression of CIDEC mRNA and protein was detected by RT-qPCR and Western blot; (4) the effect of ApoA5 on the expression of C/EBPβ mRNA and protein was detected by RT-qPCR and Western blot; (5) using lentiviral transfection technique, we overexpressed the gene of CIDEC in AMSC and cells were divided into lentiviral negative control group, lentiviral over-expressed CIDEC group and lentiviral over-expressed CIDEC+ApoA5 intervention group (the ApoA5 intervention concentration was 1 200 ng/ml). Thereby, we examined the effect of ApoA5 on the above indicators in adipogenic differentiation of AMSC in the case of CIDEC overexpression. <b>Results:</b> (1) Effect of ApoA5 on TG content in AMSC: on the 7th and 14th day after the intervention, the TG levels were lower in ApoA5 600 and 1 200 ng/ml group AMSC than those in the control group (all <i>P<</i>0.05). (2) The effect of ApoA5 on the expression of aP2 and FAS mRNA in AMSC: on the 7th day after intervention, the expression levels of aP2 and FAS mRNA were significantly lower in ApoA5 600 and 1 200 ng/ml group than those in the control group (all <i>P<</i>0.05). On the 14th day after intervention, the expression levels of aP2 and FAS mRNA were lower in ApoA5 600 and 1 200 ng/ml group than those in the control group (all <i>P<</i>0.05). (3) The effect of ApoA5 on the mRNA and protein expression of CIDEC in AMSC: on the 7th day after intervention, the mRNA and relative protein expression levels of CIDEC were significantly lower in AMSC of ApoA5 600 and 1 200 ng/ml group than those of the control group (all <i>P<</i>0.05). On the 14th day after intervention, the mRNA and relative protein levels of CIDEC were further reduced in ApoA5 600 and 1 200 ng/ml AMSC groups than those in the control group (all <i>P<</i>0.05). (4) The effect of ApoA5 on C/EBPβ mRNA and protein expression in AMSC: on the 7th day after intervention, C/EBPβ mRNA and relative protein expression levels were significantly lower in ApoA5 600 and 1 200 ng/ml group than those in the control group (all <i>P<</i>0.05). On the 14th day after intervention, the levels of C/EBPβ mRNA and relative protein were lower in ApoA5 600 and 1 200 ng/ml group than those in the control group (all <i>P<</i>0.05). (5) The effect of ApoA5 on the content of TG in AMSC after CIDEC overexpression: on the 7th and 14th day after intervention, the TG contents in AMSC were higher in the lentivirus over-expressed CIDEC group than in the lentivirus negative control group (both <i>P<</i>0.05). However, TG contents in AMSC were similar between the over-expressed CIDEC group and the CIDEC+ApoA5 over-expression group (both <i>P></i>0.05). (6) The effect of ApoA5 on the expression of aP2 and FAS mRNA in AMSC after CIDEC overexpression: on the 7th day after intervention, the expression levels of aP2 and FAS mRNA in AMSC were higher in the lentivirus over-expressed CIDEC group than in the lentivirus negative control group (both <i>P<</i>0.05). On the 14th day after intervention, the expression level of aP2 mRNA in the AMSC was higher in the lentivirus over-expressed CIDEC group than in the lentivirus negative control group (<i>P<</i>0.05). On the 7th and 14th day after intervention, the expression levels of aP2 and FAS mRNA in AMSC were similar between the lentivirus over-expressed CIDEC group and the lentivirus over-expressed CIDEC+ApoA5 group (all <i>P></i>0.05). (7) The effect of ApoA5 on the expression of C/EBPβ mRNA and protein in AMSC after CIDEC overexpression: on the 7th day after intervention, the mRNA and relative protein expressions of C/EBPβ in AMSC were higher in lentivirus-overexpressed CIDEC group than in lentivirus negative control group (both <i>P</i> <0.05). On the 14th day after intervention, C/EBPβ mRNA and protein expression levels in AMSC were higher in the lentivirus over-expressed CIDEC group than in the lentivirus negative control group (both <i>P<</i>0.05). On the 7th and 14th day after intervention, the expressions of C/EBPβ mRNA and protein in AMSC were similar between lentivirus over-expressed CIDEC group and lentivirus over-expression CIDEC+ApoA5 intervention group (all <i>P></i>0.05). <b>Conclusions:</b> ApoA5 can inhibit the adipogenic differentiation of AMSC,and this effect may be mediated by down-regulating the expression of CIDEC. Furthermore, our results indicate that CIDEC could be considered as a key factor in adipogenic differentiation.

Mesenchymal stromal/stem cells (MSCs) have shown potential in the treatment of degenerative diseases, including ischemia/reperfusion injury (IRI), which occurs during organ transplantation and represents the main cause of post-transplant graft dysfunction. However, MSCs have heterogeneous characteristics, and studies of MSCs therapy have shown a variety of outcomes. To establish a new effective MSCs therapy, we developed an adipose-derived mesenchymal stromal/stem cell line (ASCL) and compared its therapeutic effects on primary adipose-derived MSCs (ASCs) using a hepatocyte co-culture model of hypoxia/reoxygenation in vitro and a rat model of hepatic IRI in vivo. The results showed that both ASCL and ASCs protect against hypoxia by improving hepatocyte viability, inhibiting reactive oxygen species release, and upregulating transforming growth factor-β in vitro. In vivo, ASCL or ASCs were infused into the spleen 24 h before the induction of rat hepatic IRI. The results showed that ASCL significantly improved the survival outcomes compared with the control (normal saline infusion) with the significantly decreased serum levels of liver enzymes and less damage to liver tissues compared with ASCs. Both ASCL and ASCs suppressed NOD-like receptor family pyrin domain-containing 3 inflammasome activation and subsequently reduced the release of activated IL-1β and IL-18, which is considered an important mechanism underlying ASCL and ASCs infusion in hepatic IRI. In addition, ASCL can promote the release of interleukin-1 receptor antagonist, which was previously reported as a key factor in hampering the inflammatory cascade during hepatic IRI. Our results suggest ASCL as a new candidate for hepatic IRI treatment due to its relatively homogeneous characteristics.

Objective To investigate the effect of local injection of rat adipose-derived mesenchymal stem cells (ADSCs)on the phenotype of macrophages in skin wound. Methods The cryopreserved primary SD rat ADSCs were resuscitated and then sub-cultured. ADSCs of the third generation were used in the experiments. Thirty six SD rats were divided into ADSCs group (n=18) and control group(n=18) by random numbers table method. The full thickness skin wounds were established on both sides of the spine. After the model establishment 0.2 ml ADSCs suspension labeled by live cell stain Chloromethylbenzamido derivatives of 1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindo-carbocyanine perchlorate (CM-Dil) with the concentration of 5×106/ml was subcutaneously annularly injected in the skin wound of SD rats in ADSCs group. The SD rats in control group were given 0.2 ml serum-free Dulbecco modified Eagle medium (DMEM). On 3, 7, and 14 days after injury, six rats were selected from each group to measure the wound area and healing rate. The healed wound tissues were harvested to observe the morphology by HE staining. The expressions of interleukin (IL)-10 were detected by immunohistochemistry staining. The double-positive expressions of CD68 and inducible nitric oxide synthase (iNOS) (M1 type macrophages) and of CD68 and arginase-1 (Arg-1) (M2 type macrophages) were detected by immunofluorescent staining. The distribution of CM-Dil-labeled ADSCs in healed wound tissue 14 days after injury was observed by inverted fluorescence microscope. Results (1) At day 3 after injury, the wound areas in two groups were covered with crust and surrounding redness, and the wound healing rates were slightly different; at day 7 after injury, the wound area of ADSCs group was significantly smaller than that of control group, and the healing speed and rate of ADSCs group was significantly higher than that of control group (P<0.01); at day 14 after injury, the healing rate of ADSCs group was nearly 99% (P<0.01), and the healing skin tissue texture of ADSCs group was better than that of control group. (2) At day 3 after injury, there were a large number of inflammatory cells and disorganized collagen fiber in the wound areas of two groups; at day 7 after injury, the inflammatory cells infiltration reduced in ADSCs group compared with control group, and the collagen fiber arrangement in control group was in disorder; at day 14 after injury, the inflammatory cells in both groups obviously decreased, and ADSCs group had more new vessels and more orderly arrangement of collagen fiber than control group. CM Dil labeled ADSCs were seen in the healing wound tissue in ADSCs group. (3) At day 3 after injury, there was little difference in M1 type macrophage distribution in the two groups; ADSCs group had more M2 type macrophage cells than control group significantly (P<0.01); the expression of IL 10 in ADSCs group was not high, which did not differ from that of control group; at days 7 and 14 after injury, ADSCs group has fewer M1 type macrophage cells, more M2 type macrophage cells, and higher expressions of IL-10 than control group (P<0.01). Conclusion The ADSCs trasplantation can promote the change from M1 type to M2 type macrophages, facilitating wound regeneration and healing. Key words: Mesenchymal stem cells; Macrophages; Wound healing

BackgroundCanine inflammatory brain disease (IBD) is a severe inflammatory disorder characterized by infiltration of activated immune cell subsets into the brain and spinal cord. Multipotent mesenchymal stromal cells (MSCs) are a promising therapy for IBD, based on their potent pro-angiogenic, neuroprotective, and immunomodulatory properties. The aims of this study were to compare the immunomodulatory attributes of canine adipose-derived MSCs (ASCs) and placenta-derived MSCs (PMSCs) in vitro. These data will serve as potency information to help inform the optimal MSC cell source to treat naturally occurring canine IBD.MethodsIndoleamine 2,3 dioxygenase (IDO) activity and prostaglandin E2 (PGE2) concentration at baseline and after stimulation with interferon gamma (IFNγ) and/or tumor necrosis factor alpha (TNFα) were measured from canine ASC and PMSC cultures. Leukocyte suppression assays (LSAs) were performed to compare the ability of ASCs and PMSCs to inhibit activated peripheral blood mononuclear cell (PBMC) proliferation. IDO activity and PGE2; interleukin (IL)-2, IL-6, and IL-8; TNFα; and vascular endothelial growth factor (VEGF) concentrations were also measured from co-culture supernatants. Cell cycle analysis was performed to determine how ASCs and PMSCs altered lymphocyte proliferation.ResultsActivated canine MSCs from both tissue sources secreted high concentrations of IDO and PGE2, after direct stimulation with IFNγ and TNFα, or indirect stimulation by activated PBMCs. Both ASCs and PMSCs inhibited activated PBMC proliferation in LSA assays; however, PMSCs inhibited PBMC proliferation significantly more than ASCs. Blocking PGE2 and IDO in LSA assays determined that PGE2 is important only for ASC inhibition of PBMC proliferation. Activated ASCs increased IL-6 and VEGF secretion and decreased TNFα secretion, while activated PMSCs increased IL-6, IL-8, and VEGF secretion. ASCs inhibited lymphocyte proliferation via cell cycle arrest in the G0/G1 and PMSCs inhibited lymphocyte proliferation via induction of lymphocyte apoptosis.ConclusionOur results demonstrate that ASCs and PMSCs have substantial in vitro potential as a cell-based therapy for IBD; however, PMSCs more potently inhibited lymphocyte proliferation by inducing apoptosis of activated lymphocytes. These data suggest that the mechanism by which ASCs and PMSCs downregulate PBMC proliferation differs. Additional studies may elucidate additional mechanisms by which canine MSCs modulate neuroinflammatory responses.

Unraveling The Effects of DICER1 Overexpression on Immune-Related Genes Expression in Mesenchymal Stromal/Stem Cells: Insights for Therapeutic Applications.

With the emergence of coronavirus disease-2019, researchers have gained interest in the therapeutic efficacy of mesenchymal stem/stromal cells (MSCs) in acute respiratory distress syndrome; however, the mechanisms of the therapeutic effects of MSCs are unclear. We have previously reported that adipose-derived MSCs (AD-MSCs) strengthen the barrier function of the pulmonary vessels in scaffold-based bioengineered rat lungs. In this study, we evaluated whether AD-MSCs could enhance the intercellular barrier function of lung epithelial cells in vitro using a transwell coculture system. Transepithelial electrical resistance (TEER) measurements revealed that the peak TEER value was significantly higher in the AD-MSC coculture group than in the AD-MSC non-coculture group. Similarly, the permeability coefficient was significantly decreased in the AD-MSC coculture group compared to that in the AD-MSC non-coculture group. Immunostaining of insert membranes showed that zonula occuldens-1 expression was significantly high at cell junctions in the AD-MSC coculture group. Moreover, cell junction-related gene profiling showed that the expression of some claudin genes, including claudin-4, was upregulated in the AD-MSC coculture group. Taken together, these results showed that AD-MSCs enhanced the barrier function between lung epithelial cells, suggesting that both direct adhesion and indirect paracrine effects strengthened the barrier function of lung alveolar epithelium in vitro.

Transplantation of adipose-derived mesenchymal stromal/stem cells (ASCs) has successfully alleviated the severity of psoriasis. Although several therapeutic mechanisms of mesenchymal stromal/stem cells (MSCs) for psoriasis have been elucidated using the imiquimod (IMQ)-induced psoriasis-like dermatitis model, the effects of MSC transplantation on pathways other than the interleukin (IL)-23/T helper 17 (Th17) axis, including the IL-36 pathway, remain unclear. In this study, we aimed to investigate the efficacy of ASC transplantation for the IMQ-induced psoriasis-like dermatitis in male C57BL/6J mice, and to elucidate its effects on the IL-36 pathway as well as the IL23/Th17 axis. ASCs (2.0 × 106 cells) from mouse inguinal white adipose tissue were subcutaneously injected into the dorsal skin of mice. After the topical application of IMQ cream for 5 consecutive days, objective severity scores, cytokine gene expression levels, and neutrophil infiltration grade were determined to evaluate their efficacy. Anti-IL-23p19 antibody treatment was used for comparison. ASCs slightly ameliorated IMQ-induced epidermal thickening, although anti-IL-23p19 antibodies had no effect on any skin manifestations. Anti-IL-23p19 antibody and ASC suppressed the expressions of Il17a, Il17f, and Il22 mRNAs and neutrophil infiltration in IMQ-applied skin, but not the expression of Il1f6 and Il1f9. ASC also suppressed the expressions of Il23, Il6, Il1b, Tnfa, Lipocalin-2, and Cxcl5 mRNAs, which were not suppressed by anti-IL-23p19 antibody treatment. In conclusion, ASC transplantation suppressed activation of the IL-23/Th17 axis and neutrophil infiltration, and inhibited the activation of a broader range of inflammatory mediators except for IL-36 expression in IMQ-applied skin compared with anti-IL-23p19 antibody treatment.

Mesenchymal stromal cells (MSCs) have been used in the treatment of Crohn's disease (CD) because of the immunomodulatory ability. The aim of this study was to investigate the therapeutic effect of adipose-derived MSCs (AD-MSCs) and to compare the therapeutic effect of AD-MSCs with that of bone marrow MSCs (BM-MSCs) in a murine model of CD. Murine colitis model of CD was created by trinitrobenzene sulfonic acid (TNBS). Twelve hours after treatment with TNBS, the mouse model was injected with MSCs intraperitoneally. Real-time polymerase chain reaction and immunohistochemistry staining were used to measure the expression levels of inflammatory cytokines in colonic tissues to investigate the therapeutic effect of AD-MSCs. The ten-day survival was recorded after infusion of MSCs. Intraperitoneal injection of MSCs alleviated the clinical and histopathologic severity of intestinal inflammation, and improved the survival of the TNBS-induced mouse model of CD. AD-MSCs could effectively increase the expression of interleukin-10 and reduce the secretion of pro-inflammatory cytokines including tumor necrosis factor-α, interleukin-12, and vascular endothelial growth factor. The mucosal injury was repaired by AD-MSCs. These effects were comparable between AD-MSCs and BM-MSCs. The therapeutic effect appears similar between AD-MSCs and BM-MSCs in treating CD. AD-MSCs may be a potential alternative of cell-based therapy for CD.