Current Issue : January - March Volume : 2015 Issue Number : 1 Articles : 6 Articles
Introduction: Autologous mesenchymal stem cells (MSCs) are an attractive concept in regenerative medicine, but\ntheir mechanism of action remains poorly defined. No immune response is reported after in vivo injection of\nallogeneic equine MSCs or embryo-derived stem cells (ESCs) into the equine tendon, which may be due to the\ncells� immune-privileged properties. This study further investigates these properties to determine their potential for\nclinical application in other tissues.\nMethods: Mitomycin C-treated MSCs, ESCs, or differentiated ESCs (dESCs) were cultured with allogeneic equine\nperipheral blood mononuclear cells (PBMCs), and their effect on PBMC proliferation, in the presence or absence of\ninterferon-gamma (IFN-?) was determined. MSCs and super-antigen (sAg)-stimulated PBMCs were co-cultured\ndirectly or indirectly in transwells, and PBMC proliferation examined. Media from MSC culture were harvested and\nused for PBMC culture; subsequent PBMC proliferation and gene expression were evaluated and media assayed for\nIFN-?, tumor necrosis factor alpha (TNF-?), and interleukin (IL)-10 and IL-6 proteins with enzyme-linked immunosorbent\nassay (ELISA).\nResults: Co-culture of PBMCs with ESCs or dESCs did not affect baseline proliferation, whereas co-culture with\nMSCs significantly suppressed baseline proliferation. Stimulation of PBMC proliferation by using super-antigens\n(sAgs) was also suppressed by co-culture with MSCs. Inhibition was greatest with direct contact, but significant\ninhibition was produced in transwell culture and by using MSC-conditioned media, suggesting that soluble factors\nplay a role in MSC-mediated immune suppression. The MSCs constitutively secrete IL-6, even in the absence of\nco-culture with PBMCs. MSC-conditioned media also brought about a change in the cytokine-expression profile of\nsAg-stimulated PBMCs, significantly reducing PBMC expression of IL-6, IFN-?, and TNF-?.\nConclusions: Equine MSCs and ESCs possess a degree of innate immune privilege, and MSCs secrete soluble\nfactors that suppress PBMC proliferation and alter cytokine expression. These properties may make possible\nthe future clinical use of allogeneic stem cells to help standardize and broaden the scope of treatment of\ntissue injuries....
Introduction: Adipose tissue-derived stromal cells (ADSCs) are abundant and easy to obtain, but the diversity of\ndifferentiation potential from different locations may vary with the developmental origin of their mesenchymal\ncompartment. We therefore aim to compare the myogenic differentiation and reparative activity of ADSCs derived\nfrom the pericardial tissue to ADSCs of subcutaneous origin.\nMethods: Pericardial and inguinal adipose tissues from Wistar rats were surgically obtained, and the stromal\nfraction was isolated after enzymatic digestion. The phenotypic epitopes of the resultant two types of ADSCs were\nanalyzed with flow cytometry, and the expression of transcriptional factors was analyzed with immunostaining.\nFurthermore, their potential toward adipogenic, osteogenic, and myogenic differentiation also was compared.\nFinally, the reparative activity and the resultant functional benefits were examined by allograft transplantation into\nan infarcted model in rats.\nResults: ADSCs from two adipose sources showed identical morphology and growth curve at the initial stage, but\ninguinal ADSCs (ingADSCs) sustained significantly vigorous growth after 25 days of cultivation. Although both\nADSCs shared similar immunophenotypes, the pericardial ADSCs (periADSC) intrinsically exhibited partial expression\nof transcription factors for cardiogenesis (such as GATA-4, Isl-1, Nkx 2.5, and MEF-2c) and more-efficient myogenic\ndifferentiation, but less competent for adipogenic and osteogenic differentiation. After in vivo transplantation,\nperiADSCs exhibited significantly vigorous reparative activity evidenced by thickening of ventricular wall and\npronounced vasculogenesis and myogenesis, although the majority of prelabeled cells disappeared 28 days after\ntransplantation. The structural repair also translated into functional benefits of hearts after infarction.\nConclusions: Although two sources of ADSCs are phenotypically identical, pericADSCs constituted intrinsic\nproperties toward myogenesis and vasculogenesis, and thus provided more potent reparative effects after\ntransplantation; therefore, they represent an attractive candidate cell donor for cardiac therapy....
The optimal cell lineage for cardiac-regeneration approaches remains mysterious. Additionally, electrical\nstimulation promotes cardiomyogenic differentiation of stimulated cells. Therefore, we hypothesized that\nelectrical conditioning of cardiomyocyte progenitor cells (CMPCs) might enrich their cardiovascular potential.\nCMPCs were isolated from human adult atrial appendages, characterized, and electrically stimulated for 7 and\n14 days. Electrical stimulation modulated CMPCs gene and protein expression, increasing all cardiac markers.\nGATA-binding protein 4 (GATA4) early transcription factor was significantly overexpressed (P = 0.008), but also\nits coactivator myocyte enhancer factor 2A (MEF2A) was upregulated (P = 0.073) under electrical stimulation.\nMoreover, important structural proteins and calcium handling-related genes were enhanced. The cardioregeneration\ncapability of CMPCs is improved by electrical field stimulation. Consequently, short-term electrical stimulation\nshould be a valid biophysical approach to modify cardiac progenitor cells toward a cardiogenic phenotype, and\ncan be incorporated into transdifferentiation protocols. Electrostimulated CMPCs may be best-equipped cells for\nmyocardial integration after implantation....
Introduction: Our previous works demonstrated that systemic orbital fat-derived stem cell (OFSC) transplantation\nwas effective in ameliorating lipopolysaccharide (LPS)-induced extensive acute lung injury (ALI) in vivo mainly\nthrough paracrine regulation of macrophage-mediated cytokine-storm. In this study, we explore the molecular\nmechanism(s) of OFSCs regulating macrophage activity in a cytokine-inducible fashion.\nMethods: LPS (100 ng/ml)-activated macrophages were treated by conditioned medium from OFSCs (OFSCs-CM)\nor non-contact cultured with OFSCs for 6 hours. The potency of OFSCs on macrophage proliferation and\npro-inflammation ability were determined. Expression levels of pro-inflammatory cytokines in macrophages,\ninducible immuno-modulatory factors in OFSCs, were investigated. Deep sequencing analysis as well as interaction\nbetween microRNA (miRNA) and genes of immuno-modulators in OFSCs induced by activated macrophages was\npredicted by miRTar. Transfection of miRNA inhibitor into OFSCs was performed. Real-time RT-PCR and transplantation\nof OFSCs into mice with LPS-induced ALI confirmed the in vitro and in vivo mechanism.\nResults: The paracrine effect of OFSCs on inhibition of macrophage pro-inflammatory cytokine release was more\npotent than induction of macrophage G0/G1 cell cycle arrest. OFSCs-CM suppressed LPS-induced inducible nitric oxide\nsynthetase and the pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-?), interleukin (IL)-1 alpha, and\nIL-1 beta expression in macrophages. Under non-contact culture, LPS-activated macrophages effectively triggered the\nexpression of soluble immuno-modulating factors in OFSCs, i.e., IL-10, IL-1 receptor antagonist (IL-1 RA), indoleamine\n2,3-dioxygenase, and soluble TNF receptor type II (sTNF RII). Under miRTar prediction, miR-671-5p was identified as a\ncritical microRNA in regulation of multiple immune-modulating factors in OFSCs response to macrophages. The\nbaseline level of miR-671-5p was high in OFSCs, and down-regulation of miR-671-5p upon co-culture with\nactivated macrophages was observed. MiR-671-5p inhibitor transfection into OFSCs selectively enhanced the IL-1\nRA and sTNF RII expressions. In addition, inhibition of miR-671-5p in OFSCs enhanced the anti-inflammatory ability\nagainst LPS-induced ALI.\nConclusion: The paracrine effect of OFSCs inhibits the pro-inflammatory ability and proliferation of macrophages.\nThe immune-modulation capacity of OFSCs can be triggered by activated macrophages, and down-regulation of\nmiR-671-5p enhances OFSC immuno-modulation ability by up-regulating IL-1 RA and sTNF RII expression....
Introduction: Mesenchymal stromal cells (MSC) are an integral cellular component of the tumor microenvironment.\nNevertheless, very little is known about MSC originating from human malignant tissue and modulation of these\ncells by tumor-derived factors. The aim of this study was to isolate and characterize MSC from head and neck\nsquamous cell carcinoma (HNSCC) and to investigate their interaction with tumor cells.\nMethods: MSC were isolated from tumor tissues of HNSCC patients during routine oncological surgery.\nImmunophenotyping, immunofluorescence and in vitro differentiation were performed to determine whether the isolated\ncells met the consensus criteria for MSC. The cytokine profile of tumor-derived MSC was determined by enzyme-linked\nimmunosorbent assay (ELISA). Activation of MSC by tumor-conditioned media was assessed by measuring cytokine release\nand expression of CD54. The impact of MSC on tumor growth in vivo was analyzed in a HNSCC xenograft model.\nResults: Cells isolated from HNSCC tissue met the consensus criteria for MSC. Tumor-derived MSC constitutively\nproduced high amounts of interleukin (IL)-6, IL-8 and stromal cell-derived factor (SDF)-1?. HNSCC-derived factors\nactivated MSC and enhanced secretion of IL-8 and expression of CD54. Furthermore, MSC provided stromal support\nfor human HNSCC cell lines in vivo and enhanced their growth in a murine xenograft model.\nConclusions: This is the first study to isolate and characterize MSC from malignant tissues of patients with HNSCC. We\nobserved cross-talk of stromal cells and tumor cells resulting in enhanced growth of HNSCC in vivo....
Introduction: Autologous mesenchymal stem cell (MSC) injection into naturally-occurring equine tendon injuries\nhas been shown to be safe and efficacious and protocols inform translation of the technique into humans. Efficient\ntransfer of cells from the laboratory into tissue requires well-validated transport and implantation techniques.\nMethods: Cell viability in a range of media was determined over 72 hours and after injection through a 19G,\n21G or 23G needle. Viability, proliferation and apoptosis were analysed using TrypanBlue, alamarBlue�® and\nAnnexinV assays.\nResults: Cell viability was similar in all re-suspension media following 24 hour storage, however cell death was\nmost rapid in bone marrow aspirate, platelet-rich plasma and serum after longer storage. Cryogenic media exhibited\ngreatest viability regardless of storage time. Cell proliferation after 24 and 72 hour storage was similar for all media,\nexcept after 24 hours in serum wherein proliferation was enhanced. MSC tri-lineage differentiation and viability did\nnot significantly change when extruded through 19G, 21G or 23G needles, but 21G and 23G needles significantly\nincreased apoptotic cells compared to 19G and non-injected controls. All gauges induced a decrease in metabolic\nactivity immediately post-injection but cells recovered by 2 hours.\nConclusions: These data indicate storage and injection influence viability and subsequent cell behaviour and\nprovide recommendations for MSC therapy that implantation of cells should occur within 24 hours of recovery\nfrom culture, using larger needle bores....
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