Migration Of Marrow Mesenchymal Stem Cells Research Articles

Ginsenoside CK cooperates with bone mesenchymal stem cells to enhance angiogenesis post-stroke via GLUT1 and HIF-1α/VEGF pathway.

The transplantation of bone marrow mesenchymal stem cells (MSCs) in stroke is hindered by the restricted rates of survival and differentiation. Ginsenoside compound K (CK), is reported to have a neuroprotective effect and regulate energy metabolism. We applied CK to investigate if CK could promote the survival of MSCs and differentiation into brain microvascular endothelial-like cells (BMECs), thereby alleviating stroke symptoms. Therefore, transwell and middle cerebral artery occlusion (MCAO) models were used to mimic oxygen and glucose deprivation (OGD) in vitro and in vivo, respectively. Our results demonstrated that CK had a good affinity for GLUT1, which increased the expression of GLUT1 and the production of ATP, facilitated the proliferation and migration of MSCs, and activated the HIF-1α/VEGF signaling pathway to promote MSC differentiation. Moreover, CK cooperated with MSCs to protect BMECs, promote angiogenesis and vascular density, enhance neuronal and astrocytic proliferation, thereby reducing infarct volume and consequently improving neurobehavioral outcomes. These results suggest that the synergistic effects of CK and MSCs could potentially be a promising strategy for stroke.

Ultrasound-targeted microbubble destruction promotes PDGF-primed bone mesenchymal stem cell transplantation for myocardial protection in acute Myocardial Infarction in rats

BackgroundUltrasound-targeted microbubble destruction (UTMD) has emerged as a promising strategy for the targeted delivery of bone marrow mesenchymal stem cells (MSCs) to the ischemic myocardium. However, the limited migration capacity and poor survival of MSCs remains a major therapeutic barrier. The present study was performed to investigate the synergistic effect of UTMD with platelet-derived growth factor BB (PDGF-BB) on the homing of MSCs for acute myocardial infarction (AMI).MethodsMSCs from male donor rats were treated with PDGF-BB, and a novel microbubble formulation was prepared using a thin-film hydration method. In vivo, MSCs with or without PDGF-BB pretreatment were transplanted by UTMD after inducing AMI in experimental rats. The therapeutic efficacy of PDGF-BB-primed MSCs on myocardial apoptosis, angiogenesis, cardiac function and scar repair was estimated. The effects and molecular mechanisms of PDGF-BB on MSC migration and survival were explored in vitro.ResultsThe results showed that the biological effects of UTMD increased the local levels of stromal-derived factor-1 (SDF-1), which promoted the migration of transplanted MSCs to the ischemic region. Compared with UTMD alone, UTMD combined with PDGF-BB pretreatment significantly increased the cardiac homing of MSCs, which subsequently reduced myocardial apoptosis, promoted neovascularization and tissue repair, and increased cardiac function 30 days after MI. The vitro results demonstrated that PDGF-BB enhanced MSC migration and protected these cells from H2O2-induced apoptosis. Mechanistically, PDGF-BB pretreatment promoted MSC migration and inhibited H2O2-induced MSC apoptosis via activation of the phosphatidylinositol 3-kinase/serine-threonine kinase (PI3K/Akt) pathway. Furthermore, crosstalk between PDGF-BB and stromal-derived factor-1/chemokine receptor 4 (SDF-1/CXCR4) is involved in the PI3K/AKT signaling pathway.ConclusionThe present study demonstrated that UTMD combined with PDGF-BB treatment could enhance the homing ability of MSCs, thus alleviating AMI in rats. Therefore, UTMD combined with PDGF-BB pretreatment may offer exciting therapeutic opportunities for strengthening MSC therapy in ischemic diseases.

Neuritin Promotes Bone Marrow-Derived Mesenchymal Stem Cell Migration to Treat Diabetic Peripheral Neuropathy.

The purpose of this study is to explore the effect and mechanism of neuritin overexpression in the bone marrow on peripheral neuropathy in type 2 diabetic (db/db) mice. We analyzed the impact of bone marrow neuritin overexpression on diabetic peripheral neuropathy and migration of bone marrow mesenchymal stem cells in db/db mice. Antagonists were used to inhibit the stromal cell-derived factor (SDF)-1α/C-X-C chemokine receptor type 4 (CXCR4)-phosphoinositide 3-kinase (PI3K)/Akt signaling pathway in primary cultured bone marrow mesenchymal stem cells. Immunofluorescence, transmission electron microscopy, Oil Red O staining, and transwell migration assays were used. Bone marrow-specific overexpression of neuritin in db/db mice was successfully established. Overexpression of neuritin in the bone marrow ameliorated hyperglycemia, prevented diabetic peripheral neuropathy, protected the ultrastructure of the sciatic nerve and intra-epidermal nerve fiber density, and promoted Schwann cell proliferation and remyelination in the sciatic nerve. Moreover, it ameliorated fat accumulation, adipocyte number, and vascular and nerve densities; decreased glutamate content in serum and bone marrow; restored gradient SDF-1α contents between bone marrow, blood, and sciatic nerve; and promoted impaired diabetic bone marrow mesenchymal stem cell migration. Neuritin improves bone marrow mesenchymal stem cell migration via the SDF-1α/CXCR4-PI3K/Akt signaling pathway in vitro. Overexpression of neuritin in the bone marrow can locally ameliorate neuropathy in the bone marrow. This improves the migration capability of bone marrow mesenchymal stem cells and repairs diabetic peripheral neuropathy, at least partly by activating the PI3K/Akt pathway through the SDF-1α/CXCR4 axis.

Chemical Compounds Released from Specific Osteoinductive Bioactive Materials Stimulate Human Bone Marrow Mesenchymal Stem Cell Migration.

In this work, a poly(L-lactide-co-glycolide) (PLGA)-based composite was enriched with one of the following sol-gel bioactive glasses (SBG) at 50 wt.%: A1—40 mol% SiO2, 60 mol% CaO, CaO/SiO2 ratio of 1.50; S1—80 mol% SiO2, 20 mol% CaO, CaO/SiO2 ratio of 0.25; A2—40 mol% SiO2, 54 mol% CaO, 6 mol% P2O5, CaO/SiO2 ratio of 1.35; S2—80 mol% SiO2,16 mol% CaO, 4 mol% P2O5, CaO/SiO2 ratio of 0.20. The composites and PLGA control sheets were then soaked for 24 h in culture media, and the obtained condition media (CM) were used to treat human bone marrow stromal cells (hBMSCs) for 72 h. All CMs from the composites increased ERK 1/2 activity vs. the control PLGA CM. However, expressions of cell migration-related c-Fos, osteopontin, matrix metalloproteinase-2, C-X-C chemokine receptor type 4, vascular endothelial growth factor, and bone morphogenetic protein 2 were significantly increased only in cells treated with the CM from the A1/PLGA composite. This CM also significantly increased the rate of human BMSC migration but did not affect cell metabolic activity. These results indicate important biological markers that are upregulated by products released from the bioactive composites of a specific chemical composition, which may eventually prompt osteoprogenitor cells to colonize the bioactive material and accelerate the process of tissue regeneration.

Molecular Mechanism of Neurotrophic Factor-Activated Long Non-Coding RNA Plasmacytoma Variant Translocation 1 Promoting Mesenchymal Stem Cell Migration and Repair of Fractures

It has been reported that neurotrophic factor (NF) promotes bone marrow mesenchymal stem cells (MSCs) migration to repair fractures. However, whether and how lncRNA PVT1 regulates differentiation induced by neurotrophic factors to promote MSC migration to repair fractures has not been explored. To explore the molecular mechanism of neurotrophic factor activating lncRNA PVT1 to promote MSC migration and repair fractures. Differential expression of neurotrophic factors stimulated by MSCs was analyzed based on microarray lncRNA and lncRNAs was further verified by qRT-PCR. The conditions of promoting MSC migration and osteogenic differentiation were identified by trans-fection of lncRNA PVT1 overexpressed plasmids and inhibitor and the targets of its regulation were confirmed by target gene prediction tools. In this study lncRNA array and qRT-PCR showed that lncRNA PVT1 was significantly down-regulated during neurotrophic factor-induced MSCs differentiation. Transfection of lncRNA PVT1 overexpression plasmid significantly inhibited the expression of osteogenic markers alkaline phosphatase (ALP) and osteopontin (OPN) in MSCs, while transfection of lncRNA PVT1 inhibitor promoted the expression of alkaline phosphatase (ALP) and osteopontin (OPN). lncRNA PVT1 is a negative regulator of MSCs differentiation induced by neurotrophic factors. The distal deletion homologous box 5(DLX5) was identified as the target of lncRNA PVT1 and the relationship between lncRNA PVT1 inhibiting the expression of DLX5 and the osteogenic differentiation of MSCs was verified in MSCs. lncRNA PVT1 negatively regulates the migration and differentiation of MSCs induced by neurotrophic factors by targeting DLX5, providing the foundation for bone repair.

17β-estradiol promotes bone marrow mesenchymal stem cell migration mediated by chemokine upregulation

Bone marrow-derived cells engraft to the uterine endometrium and contribute to endometriosis. This study sought to further confirm that estrogen can promote the migration of bone marrow mesenchymal stem cells (BMSCs) and to investigate the function of estrogen on the secretion of chemokines during BMSC migration. BMSCs were treated with or without 17β-estradiol, cultured with or without endometrial stromal cells (ESCs), or pretreated with or without AMD 3100 (an antagonist of the SDF-1α receptor) before co-culture. A migration assay was used to investigate the changes in the migration of BMSCs. The secretion of chemokines in the co-culture medium was detected by chemokine analysis, and the mRNA expression of SDF-1α in cells was tested using quantitative real-time PCR. The results revealed that the migration of BMSCs was promoted by ESC, and the migration ability of BMSCs was enhanced after treatment with 17β-estradiol (p < 0.05). Through chemokine analysis, we further showed that 17β-estradiol promoted the secretion of chemokines especially for SDF-1α (p < 0.05). Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that these chemokines were mainly linked to the cytokine signaling pathway and interaction with cytokines receptors. Furthermore, the expression of SDF-1α mRNA was significantly increased in the 17β-estradiol treatment group (p < 0.001), and the migration of BMSCs was blocked by the use of our SDF-1α antagonist (p < 0.01). Our results indicate that 17β-estradiol could promote the chemotaxis and migration of BMSCs by up-regulating the secretion of chemokines, especially SDF-1α. Our study provides additional evidence to support and supplement the stem cell theory of endometriosis.

Construction of chemokine substance P-embedded biomimetic multilayer onto bioactive magnesium silicate-titanium implant for bone regeneration

Stem cell-mediated biological processes play very important roles in bone regeneration and tissue repair. It's urgent to design new generations of tissue inductive implantable biomaterials that can regulate the biological behavior of autologous bone marrow mesenchymal stem cells (MSCs) in situ. Therefore, we designed and constructed chemokine substance P (SP)-intercalated multilayer films containing chitosan (Chi) /gelatin (Gel), which were used to cover the surface of alkaline magnesium silicate (MgSiO3)-titanium (Ti). The MgSi/LBL-SP substrate could regulate the release of SP, Mg and Si from the multilayer system as well as the resultant pH changes in a mild manner to improve the biological performance. It was demonstrated that the SP released from MgSi/LBL-SP substrate could effectively enhance MSCs recruitment in vitro and in vivo through transwell assay of MSCs migration, detection of MMP2 secretion and double immunofluorescence staining of CD29+/CD90+ markers. Meanwhile, MgSi/LBL-SP group also significantly improved the proliferation and differentiation of MSCs, which were reflected by the results of MTT assay, ALP activity, collagen secretion, ECM mineralization and western blotting. On animal models, the MgSi/LBL-SP implants displayed the highest stimulation effect on cell proliferation activity and apoptosis inhibition. Moreover, the results of histological staining and micro-CT further verified that MgSi/LBL-SP implants were more beneficial for facilitating new bone formation after implantation for 1 month. Hence, this dually optimized Ti-based implant may serve as an ideal substitute material for bone tissue repair and regeneration.

Mesenchymal Stem Cell-Platelet Aggregates Increased in the Peripheral Blood of Patients with Acute Myocardial Infarction and Might Depend on the Stromal Cell-Derived Factor 1/CXCR4 Axis.

Bone marrow mesenchymal stem cells (MSCs) are a rare subset of nonhematopoietic progenitor cells and are appealing biomaterial for multiple tissue damage repairs. Transplantation of MSCs is proved to improve heart function after myocardial ischemia. However, the limitations of MSC injection approaches are equally obvious. As a multiple-function cell, platelets (PLTs) are also known playing important roles in cardiac recovery after myocardial infarction. In this study, we analyzed circulating MSC-PLT aggregate numbers in acute myocardial infarction (AMI) patients by flow cytometry. We found more MSC-PLT aggregates in patients with AMI than in healthy controls, and the patients with higher MSC-PLT aggregates had better prognosis. When stromal cell-derived factor 1 (SDF-1) binds to its receptor CXC chemokine receptor 4 (CXCR4), they play an important role in MSC migration and engraftment. We explored SDF-1 and CXCR4 expression on PLT surface by flow cytometry and found relative mean fluorescence intensity of PLT CXCR4 and the number of MSC-PLT aggregates showed a significant correlation. Meanwhile, in vitro experiments demonstrated that SDF-1/CXCR4 was crucial in MSC-PLT aggregate formation, which might suggest a novel mechanism that SDF-1/CXCR4 is involved in MSCs homing and myocardial repair after AMI. There may be another strategy to encourage myocardial repair in AMI patients by increasing the expression of SDF-1 on MSCs and promoting the formation of MSC-PLT aggregates.

Intravenous Administration of Granulocyte-Colony Stimulating Factor for Stem Cells Mobilization and Third Degree Burn Wound Healing in Rats

Introduction: Burns are one the most common skin damages which require medical intervention to be fully-recovered. In this light, tissue engineering field presents a vide verity of strategies including both scaffold-based and cell-based approaches to recover the damaged site. Materials and Methods: In this study, the effects of granulocyte-colony stimulating factor (G-CSF) administration on mobilization of the bone marrow mesenchymal stem cells (MSCs) into defect area and treatment of the skin burn wound was examined in vivo. The G-CSF was injected intravenously into rats subjected to third degree burn wound. At days 3, 5, 7, 15 and 30 post-injections, the defect site was removed and investigated by H&E and Malory’s trichrome staining. The number of MSCs in blood samples was also determined by flow cytometry assay. Results: According to the results, intravenously administration of G-CSF significantly increased collagenesis and number of fibroblast cells infiltrated into the burned site, while decreased the severity of acute inflammatory response and amount of inflammatory cells comparing to control. The number of MSCs in bloodstream, representing the rate of MSCs migration, showed a 4-fold increase in the experimental group compared to control. Conclusions: The current study suggests the potential of intravenously administration of G-CSF as an effective strategy for treatment of severe burn injuries.

Neuregulin‑1 impacting bone marrow mesenchymal stem cell migration is conducive to functional recovery following spinal cord injury.

The present study was designed to investigate the effect of neuregulin-1 (NRG1) on the migration of rat bone marrow mesenchymal stem cells (BMSCs) and evaluate the role of NRG1 in the functional recovery following spinal cord injury (SCI). Firstly, the effect of NRG1 on the mRNA expression of Snail in the BMSCs was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis; secondly, the BMSCs were transfected with a Snail-overexpression plasmid (pBabe-puro-Snail) and the expression levels of Snail and matrix metalloptoreinase-2 (MMP-2) were detected by RT-qPCR and western blot analyses; thirdly, the cell proliferation and migration of BMSCs modified with pBabe-puro-Snail were detected by methyl thiazolyl tetrazolium and migration assays, respectively; finally, functional recovery of SCI was assessed using the Basso, Beattie, and Bresnahan rating scales. The results showed that NRG1 concentration-dependently promoted the expression of Snail with a peak at 40 ng/ml and 48 h; NRG1 enhanced the promoting effect of Snail on the expression of MMP-2; the overexpression of Snail did not enhance the cell growth of the BMSCs. The NRG1-modified BMSCs promoted the functional recovery of SCI. These results suggested that NRG1 significantly promoted the expression of MMP-2 by upregulating the expression of Snail, and enhanced cell migration of the BMSCs conducive to the functional recovery of SCI.

CoCl2, a mimic of hypoxia, enhances bone marrow mesenchymal stem cells migration and osteogenic differentiation via STAT3 signaling pathway

Mesenchymal stem cells homing and migration is a crucial step during bone fracture healing. Hypoxic environment in fracture site induces bone marrow mesenchymal stem cells (BMSCs) migration, but its mechanism remains unclear. Our previous study and studies by other groups have reported the involvement of signal transducer and activator of transcription 3 (STAT3) pathway in cell migration. However, the role of STAT3 pathway in hypoxia-induced cell migration is still unknown. In this study, we investigated the role of STAT3 signaling in hypoxia-induced BMSCs migration and osteogenic differentiation. BMSCs isolated from C57BL/6 male mice were cultured in the presence of cobalt chloride (CoCl2 ) to simulate intracellular hypoxia. Hypoxia enhanced BMSCs migration, and upregulated cell migration related gene expression, that is, metalloproteinase (MMP) 7, MMP9, and C-X-C motif chemokine receptor 4. Hypoxia enhanced the phosphorylation of STAT3, and cell migration related proteins: c-jun n-terminal kinase (JNK), focal of adhesion kinase (FAK), extracellular regulated protein kinases, and protein kinase B 1/2 (ERK1/2). Moreover, hypoxia enhanced expression of osteogenic differentiation marker. Inhibition of STAT3 suppressed the hypoxia-induced BMSCs migration, cell migration related signaling molecules phosphorylation, and osteogenic differentiation related gene expression. In conclusion, our result indicates that hypoxia-induced BMSCs migration and osteogenic differentiation is via STAT3 phosphorylation and involves the cooperative activity of the JNK, FAK, and MMP9 signaling pathways.

(*)AG490 could suppress bone marrow mesenchymal stem cells migration, mineralization and bone defect healing via inhibiting Jak2-STAT3 pathway

Objective: To study the effect of Jak2-STAT3 pathway on cell proliferation, migration, mineralization and bone defect healing via simulating Jak2-STAT3 pathway inhibitor AG490 to bone marrow mesenchymal stem cells (BMSC) and bone defect mice models. Methods: The effect of AG490 on BMSC proliferation was measured by MTT (methyl thiazolyl tetrazolium) assay. Regulation of AG490 on BMSC migration was tested by scratch assay and transwell assay. The BMSC migration related gene, matrix metalloproteinase (MMP)-7, MMP-9 and CXC subfamily receptor 4 (CXCR4), regulated by AG490 was studied by real-time PCR. Western blotting was adopted to analyze the regulation of Jak2-STAT3 phosphorylation through the simulation of AG490. The alizarin red staining and alkaline phosphatase (ALP) activity assay were performed to measure the effect of AG490 on BMSC mineralization and osteogenic differentiation. Mice femur bone defect models were built to analyzed the effect of AG490 on bone remodeling. Results: AG490 significantly suppressed the migration rate of BMSC at 1 d and 2 d in the experiment group [(12.42±7.50) %, (41.8±2.6)%] compared with the control group [(55.5±9.9)%, (86.9±8.7)%] in scratch assay (P=0.000, P=0.000), the number of migrated BMSC in the experiment group (22.8± 5.9) was significantly suppressed compared with the control group (58.3±6.6) in Transwell assay (P=0.000). The expression of MMP-7, MMP-9 and CXCR4 were significantly downregulated in experiment group [(0.5± 0.1), (0.1±0.1) and (0.35±0.07)] compared with the control group [(1.1±0.1), (1.06±0.33), (1.08±0.13)] (P= 0.0003, P=0.000 and P=0.000). Also, the phosphorylation of Jak2-STAT3 was downregulated by AG490 in western blotting. After BMSCs were osteogenic induced for 14 days, the formation of mineralized nodule and the ALP activity of BMSC is significantly suppressed in experiment group (8.0±2.1) compared with the control group (35.7 ± 1.8) (P=0.0005). AG490 suppressed the bone defects healing,the expression level of phosphorylated Jak2 and phosphorylated STAT3, and the number of alkaline phosphatase positive cell at defect area in vivo is lower in experiment group than the control group. AG490 suppressed the relatively bone density at the defect area significantly (P=0.0004) at 5(th) week after the surgery. Conclusions: AG490 could suppress proliferation, migration and mineralization to of BMSCs regulate bone defect healing via inhibiting Jak2-STAT3 pathway.

Transplantation with hypoxia-preconditioned mesenchymal stem cells suppresses brain injury caused by cardiac arrest-induced global cerebral ischemia in rats.

Cardiac arrest-induced global cerebral ischemia is a main cause of neurological dysfunction in emergency medicine. Transplantation with bone marrow mesenchymal stem cells (MSCs) has been used in stroke models to repair the ischemic brain injury, but it is little studied in models with global cerebral ischemia. In the present study, a hypoxia precondition was used to improve the efficacy of MSC transplantation, given the low survival and migration rates and limited differentiation capacities of MSCs. We found that hypoxia can increase the expansion and migration of MSCs by activating the PI3K/AKT and hypoxia-inducible factor-1α/CXC chemokine receptor-4 pathways. By using a cardiac arrest-induced global cerebral ischemic model in rats, we found that transplantation of hypoxia-preconditioned MSCs promoted the migration and integration of MSCs and decreased neuronal death and inflammation in the ischemic cortex. © 2017 Wiley Periodicals, Inc.

The mechanically activated p38/MMP-2 signaling pathway promotes bone marrow mesenchymal stem cell migration in rats

ObjectiveThe aim of the present study was to investigate the effect of static strain on bone marrow mesenchymal stem cell (BMMSC) migration and whether the p38/matrix metalloproteinase-2 (MMP-2) axis plays a role in induction of BMMSC migration under mechanical strain. DesignBoth in vivo and in vitro investigations were performed. Twelve adult male Sprague-Dawley rats were randomly divided into 2 groups (n=6 per group). Rats in the experimental group underwent right mandibular distraction osteogenesis, whereas rats in the control group were subjected to osteotomy in the mandible without distraction. Immunohistochemistry and immunofluorescence were performed to evaluate phospho-p38 (p-p38) and Nestin expression. BMMSCs were isolated from rat mandibles. BMMSCs in the experimental group were subjected to static mechanical strain for 2h, whereas those in the control group underwent no strain. The biological roles of static strain and the p38/MMP-2 axis in BMMSC migration were evaluated by Transwell assays and western blotting by inhibiting p38 phosphorylation. ResultsThere were significantly more Nestin+ cells in the bone calluses of the experimental group than in those of the control group. In addition, Nestin+/p-p38+ cell numbers were significantly higher in the experimental group than in the control group, indicating that static strain activated p38 signaling in BMMSCs in vivo. In accordance with in vivo results, static strain in vitro stimulated phosphorylation of p38 in BMMSCs. Furthermore, expression of MMP-2 was elevated in BMMSCs under static strain compared with the control, and strain-induced MMP-2 expression was abolished by inhibition of p38 phosphorylation in BMMSCs. Moreover, Transwell assay results showed that static strain promoted BMMSC migration, which was abolished by inhibition of p38 phosphorylation. ConclusionsThe present study demonstrated that static strain can promote the migration ability of BMMSCs via p38/MMP-2 signaling. To the best of our knowledge, this study is the first report demonstrating that the p38/MMP-2 axis governs BMMSC migration under static mechanical strain.

Hypoxia and low-dose inflammatory stimulus synergistically enhance bone marrow mesenchymal stem cell migration.

Cell migration is necessary for numerous physiological cell processes. Although either inflammatory or hypoxic stimuli of certain dose and duration have positive influence on cell migration, their combination has not been shown to result in a synergistic effect. In this study, we investigated combined effects of hypoxia and low-dose inflammatory stimulus (one-tenth of that of a previously used concentration) on migration of human bone marrow-derived mesenchymal stem cells (BMMSCs). Our results from real-time PCR, Western blot analysis and an immunofluorescence assay, showed that dual stimulation up-regulated CXCR4 expression. Based on tablet scratch experimentation and transwell assay, the dual stimuli exhibited greater positive effects on cell migration than a single inflammatory or hypoxic stimulus. When effects of various pre-treatments on cell proliferation, differentiation and immunosuppression were screened, cells subjected to the hypoxic stimulus or dual stimuli had increased cell proliferation, while short-term inflammatory stimulus and/or hypoxic stimulus had no negative effect on cell differentiation and immunosuppression. These findings suggest that the combination of hypoxia and low-dose inflammatory stimuli enhances the potential of BMMSCs to migrate, thus identifying cell pre-treatment conditions that could enhance future stem cell-based therapeutics.

Abstract 355: Heart Failure Impairs Bone Marrow Mesenchymal Stem Cell Renewing Capacity and Migration

Background: Clinical trials have shown a sustained beneficial effect of bone marrow mesenchymal stem cells (MSCs) as a therapy for acute and chronic heart failure (HF). Clinical grade cell manufacturing of autologous and allogeneic MSCs is becoming a standard procedure. This study investigates the differences of bone marrow MSC isolation and expansion in favorable in vitro conditions. We compared MSC production from both healthy young donors and chronic HF patients. Methods: We analyzed MSC manufacturing records from five clinical trials: TAC-HFT (Ischemic cardiomyopathy (CMP), patient and donors), POSEIDON (Ischemic CMP, donors), POSEIDON DCM (Dilated CMP, donors), TRIDENT (Ischemic CMP, donors), and CRATUS (Frailty, donors). Results: All cells expressed CD105, CD90 and lacked hematopoietic marker CD45. The age of healthy donors (25.5 ± 0.7 y.o., N=24) and HF patients (56.1 ± 2.5, y.o., N=23), was significantly different (P&lt;0.0001). Collectively, the number of mononuclear cells (MNCs) isolated from bone marrow (volume range 58-125ml) didn’t differ between the groups. However, plated MNCs from healthy adults generated more MSCs than MNCs from HF patients (p0: 5.9x10^6±0.5x10^6, N=23 vs 3.8x10^6±0.4x10^6, N=24, respectively, P=0.003 and p1: 15.4x10^6±2.5x10^6, N=23 vs 10.8x10^6±1.1x10^6, N=24, respectively, P=0.036). No correlation was found between stem cell growth and age (35 to 78y.o.). Left ventricular ejection fraction (LVEF) in patients with HF had a direct correlation with MSC growth rate (P=0.03), particularly, in patients with severely depressed LVEF (&lt;30%), which had a tendency to generate less MSCs overall. Moreover, MSCs from HF patients demonstrated less migration compared to MSCs from healthy donors at 6, 10 and 24 hours (h) relative to baseline (6 h: 9.5±3.0% vs 30.7±2.1%; 10 h: 19.9±13.7% vs 53.1±2.5% and 24 h: 41.5±15.8% vs 88.9±1.6%, N=4, respectively. P=0.03). Conclusions: Despite equivalent numbers of MNCs, healthy young donors generate significantly more MSCs than HF patients, due to increased growth rate and higher number of resident stromal bone marrow stem cells. MSC migration was impaired in HF patients compared to healthy donors. HF appears to be an independent factor for MSC renewal capacity and culture phenotype.

Stromal cell-derived factor-1-directed bone marrow mesenchymal stem cell migration in response to inflammatory and/or hypoxic stimuli

ABSTRACTDirecting cell trafficking toward a target site of interest is critical for advancing stem cell therapy in clinical theranostic applications. In this study, we investigated the effects of inflammatory and/or hypoxic stimuli on the migration of bone marrow mesenchymal stem cells (BMMSCs) during in vitro culture and after in vivo implantation. Using tablet scratch experiments and observations from a transwell system, we found that both inflammatory and hypoxic stimuli significantly enhanced cell migration. However, the combination of inflammatory and hypoxic stimuli did not result in a synergistic effect. The presence of stromal cell-derived factor-1 (SDF-1) significantly enhanced cell migration irrespective of the incubation conditions, and these positive effects could be blocked by treatment with AMD3100. Based on a time course experiment, we found that preconditioning cells with either inflammatory or hypoxic stimuli for 24 h or with both stimuli for 12 h led to high levels of chemokine receptor type 4 (CXCR4) expression. In vivo studies further demonstrated that pretreatment of BMMSCs with inflammatory and/or hypoxic stimuli resulted in an increased number of systemically injected cells migrating toward skin injuries, and local SDF-1 administration significantly increased cell migration. These findings suggest that in vitro control of either inflammatory or hypoxic stimuli has significant potential to enhance SDF-1-directed BMMSC migration via the upregulation of CXCR4 expression. Although combining the stimuli did not necessarily lead to a synergistic effect, the potential to reduce the dose and time required for cell preconditioning indicates that combinations of various strategies warrant further exploration.

GW25-e2345 MiR-211 mediated hypoxia induce bone marrow mesenchymal stem cells migration through STAT5A

GW25-e2345 MiR-211 mediated hypoxia induce bone marrow mesenchymal stem cells migration through STAT5A

Sympathetic denervation-induced MSC mobilization in distraction osteogenesis associates with inhibition of MSC migration and osteogenesis by norepinephrine/adrb3.

The sympathetic nervous system regulates bone formation and resorption under physiological conditions. However, it is still unclear how the sympathetic nerves affect stem cell migration and differentiation in bone regeneration. Distraction osteogenesis is an ideal model of bone regeneration due to its special nature as a self-engineering tissue. In this study, a rat model of mandibular distraction osteogenesis with transection of cervical sympathetic trunk was used to demonstrate that sympathetic denervation can deplete norepinephrine (NE) in distraction-induced bone callus, down-regulate β3-adrenergic receptor (adrb3) in bone marrow mesenchymal stem cells (MSCs), and promote MSC migration from perivascular regions to bone-forming units. An in vitro Transwell assay was here used to demonstrate that NE can inhibit stroma-derived factor-1 (SDF-1)-induced MSC migration and expression of the migration-related gene matrix metalloproteinase-2 (MMP-2) and downregulate that of the anti-migration gene tissue inhibitor of metalloproteinase-3 (TIMP-3). Knockdown of adrb3 using siRNA abolishes inhibition of MSC migration. An in vitro osteogenic assay was used to show that NE can inhibit the formation of MSC bone nodules and expression of the osteogenic marker genes alkaline phosphatase (ALP), osteocalcin (OCN), and runt-related transcription factor-2 (RUNX2), but knockdown of adrb3 by siRNA can abolish such inhibition of the osteogenic differentiation of MSCs. It is here concluded that sympathetic denervation-induced MSC mobilization in rat mandibular distraction osteogenesis is associated with inhibition of MSC migration and osteogenic differentiation by NE/adrb3 in vitro. These findings may facilitate understanding of the relationship of MSC mobilization and sympathetic nervous system across a wide spectrum of tissue regeneration processes.

Preconditioning of Mesenchymal Stem Cells by Sevoflurane to Improve Their Therapeutic Potential

BackgroundBone marrow mesenchymal stem cells (MSCs) have been found to produce beneficial effects on ischemia-reperfusion injury. However, most of the MSCs died when transplanted into the ischemic tissue, which severely limit their therapeutic potential.MethodsUsing an in vitro model of hypoxia and serum deprivation (H/SD), we investigated the hypothesis that sevoflurane preconditioning could protect MSCs against H/SD-induced apoptosis and improve their migration, proliferation, and therapeutic potential. The H/SD of MSCs and neuron-like PC12 cells were incubated in a serum-free medium and an oxygen concentration below 0.1% for 24 h. Sevoflurane preconditioning was performed through a 2-h incubation of MSCs in an airtight chamber filled with 2 vol% sevoflurane. Apoptosis of MSCs or neuron-like PC12 cells was assessed using Annexin V-FITC/propidium iodide (PI). Furthermore, the mitochondrial membrane potential was assessed using lipophilic cationic probe. The proliferation rate was evaluated through cell cycle analysis. Finally, HIF-1α, HIF-2α, VEGF and p-Akt/Akt levels were measured by western blot.ResultsSevoflurane preconditioning minimized the MSCs apoptosis and loss of mitochondrial membrane potential. Furthermore, it increased the migration and expression of HIF-1α, HIF-2α, VEGF, and p-Akt/Akt, reduced by H/SD. In addition, neuron-like PC12 cells were more resistant to H/SD-induced apoptosis when they were co-cultured with sevoflurane preconditioning MSCs.ConclusionThese findings suggest that sevoflurane preconditioning produces protective effects on survival and migration of MSCs against H/SD, as well as improving the therapeutic potential of MSCs. These beneficial effects might be mediated at least in part by upregulating HIF-1α, HIF-2α, VEGF, and p-Akt/Akt.