Wharton's Jelly Mesenchymal Stem Cells Research Articles

Characterization and plasticity of wharton's jelly mesenchymal stem cells of goat

Mesenchymal stem cells (MSCs), obtained from several anatomical sites, have already been described, characterized and used in therapeutic models for tissue repair. The umbilical cord mesenchymal stem cells, represented by cells from arteries and veins walls, as well as Wharton's jelly are easy to be obtained, highly available, require no invasive procedure, do not present risk to donors and do not present ethical limitation. The aim of this research was to analyze the plasticity of Wharton's jelly mesenchymal stem cells (WJ-MSCs) of goat, evaluating their behavior in vitro and characterizing them immunophenotypically. Thus, tests were performed on colony forming units, viability and cell growth curve, flow cytometry analysis and plasticity potential. Goat umbilical cord matrix cells exhibited fibroblastoid morphology with colony formation and self-renewal ability, always maintaining their undifferentiated state up to the eighth passage (P8). The growth curve kinetics exhibited the LAG, LOG, and DECAY phases, without displaying a PLATEAU phase. The plasticity assay demonstrated positive differentiation for osteogenic, adipogenic and chondrogenic lines, characterized by the synthesis of intracytoplasmic granules or extracellular matrix with the presence of calcium, lipids and proteoglycans. Flow cytometry demonstrated the expression of CD90 and CD105; absence of CD14 expression. It is concluded that the cell population isolated from the Wharton's jelly of goat constitutes a representative sample of mesenchymal stem cells, with great possibilities in the field of regenerative and reproductive medicine.

Muscle functional recovery is driven by extracellular vesicles combined with muscle extracellular matrix in a volumetric muscle loss murine model

Biological scaffolds derived from decellularized tissues are being investigated as a promising approach to repair volumetric muscle losses (VML). Indeed, extracellular matrix (ECM) from decellularized tissues is highly biocompatible and mimics the original tissue. However, the development of fibrosis and the muscle stiffness still represents a major problem. Intercellular signals mediating tissue repair are conveyed via extracellular vesicles (EVs), biologically active nanoparticles secreted by the cells. This work aimed at using muscle ECM and human EVs derived from Wharton Jelly mesenchymal stromal cells (MSC EVs) to boost tissue regeneration in a VML murine model. Mice transplanted with muscle ECM and treated with PBS or MSC EVs were analyzed after 7 and 30 days. Flow cytometry, tissue analysis, qRT-PCR and physiology test were performed. We demonstrated that angiogenesis and myogenesis were enhanced while fibrosis was reduced after EV treatment. Moreover, the inflammation was directed toward tissue repair. M2-like, pro-regenerative macrophages were significantly increased in the MSC EVs treated group compared to control. Strikingly, the histological improvements were associated with enhanced functional recovery. These results suggest that human MSC EVs can be a naturally-derived boost able to ameliorate the efficacy of tissue-specific ECM in muscle regeneration up to the restored tissue function.

The effect of vascular complications of diabetes mellitus on human umbilical cord tissue and the number of Wharton Jelly's mesenchymal stem cells.

The current study investigated the change in umbilical cord tissue and the number of markers of Wharton's jelly mesenchymal stem cells (WJ-MSC) in pregnant women with gestational diabetes (GDM), with chronic diabetes who developed nephropathy as vascular complication (VC-PGDM), and healthy pregnant women as the control. The umbilical cords (UC) were investigated by the histomorphological method and the number of WJ-MSC were detected by flow-cytometry using the CD90, CD44, CD105, and CD73 markers in Wharton's jelly (WJ) isolated from fresh umbilical cords. The number of positive cells for CD 90, CD44, CD105, and CD73 were found to be elevated in the GDM group, whereas it was significantly diminished in the VC-PGDM group (p = 0.001, p = 0.001, p = 0.001, and p = 0.001). The only histopathological sign in the GDM group were an increased number of pores in the Wharton jelly. Artery wall thickness/cord diamater ratio was increased, which indicates an increase of the artery wall thickness in the VC- PGDM group (p = 0.039 and p = 0.048). The increase in umbilical cord diameter and number of Wharton jelly mesenchymal stem cells in babies of gestational diabetic mothers was considered as an effect of macrosomia seen in babies of mothers with gestational diabetes. Vasculopathy, a long-term complication of diabetes, is known to affect all tissues by causing marked lower perfusion and hypoxia, as well as a decrease in the MSC number in our study.

Evolution of left ventricular function after Wharton's jelly mesenchymal stem cells transcoronary administration: 5-year follow up in a pilot cohort of CIRCULATE-AMI Randomized Trial

Abstract Introduction CIRCULATE-Acute Myocardial Infarction is a double-blind controlled trial randomizing (RCT) in 105 consecutive patients with their first, large AMI (cMRI-LVEF ≤45% and/or cMRI-infarct size ≥10% of LV) with successful infarct-related artery (IRA) primary percutaneous coronary intervention (pPCI) to transcoronary administration of Wharton's Jelly Mesenchymal Stem Cells (WJMSCs) vs. placebo (2:1). The pilot study cohort (PSC) preceded the RCT. Aim To evaluate WJMSCs long-term safety, and evolution of left-ventricular (LV) function in CIRCULATE-AMI PSC. Material and methods 30 000 000 WJMSCs (50% labelled with 99mTc-exametazime) were administered via IRA in a ten-patient PCS (age 32–65 years, peak hs-Troponin T 17.3±9.1ng/mL and peak CK-MB 533±89U/L, cMRI-LVEF 40.3±2.7% and infarct size 20.1±2.8%) at ≈5–7 days after AMI using a cell delivery-dedicated, coronary-non-occlusive method. Other treatments were per guidelines. WJMSCs showed an unprecedented high myocardial uptake (30.2±5.3%; 95% CI 26.9–33.5%), corresponding to ≈9×10 000 000 cells retention in the infarct zone – in absence of epicardial flow or myocardial perfusion impairment (TIMI-3 in all; cTFC 45±8 vs. 44±9, p=0.51) or any hs-Troponin T elevation. Five-year follow up included cardiac Magnetic Resonance Imaging (cMRI) (at baseline, 1 year and 3 years) and detailed echocardiography (echo) at baseline, 1 year, 3 years and 5 years. Results By 5 years, one patient died from a new, non-index territory AMI. There were no other cardiovascular events and MACCE that might be related to WJMSCs transplantation. On echo (Fig), there was an increase in left ventricular ejection fraction (LVEF) between WJMSCs administration point and 1 year (37.7±2.9% vs. 48.3±2.5%, p=0.002) that was sustained at 3 years (47.2±2.6%, p=0.005 vs. baseline) and at 5 years: (44.7±3.2%, p=0.039 vs. baseline). LVEF reached a peak at 1 year after the AMI and WJMSCs transfer (Fig). cMRI data (obtained up to 3 years; 1 year 41.9±2.6% vs. 51.0±3.3%, p<0.01; 3 years 52.2±4.0%, p<0.01 vs. baseline) were consistent with the echo LVEF assessment. Conclusions 5-year follow up in CIRCULATE-AMI PSC indicates that WJMSC transcoronary application is safe and may be associated with an LVEF improvement. The magnitude of LV increase appears to peak at 1 year, suggesting a potential role for repeated WJMSCs administration(s). Currently running double-blind RCT will provide placebo-controlled insights into the WJMSCs effect(s) on changes in LV function, remodelling, scar reduction and clinical outcomes. Echo-LVEF evolution Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): STRATEGMED 265761 “CIRCULATE” National Centre for Research and Development/Poland/ZDS/00564 Jagiellonian University Medical College

Topical Instillation of Resveratrol Preconditioned Wharton’s Jelly Mesenchymal Stem Cell Secretome Preserves Ocular Surface in Experimental Model of Severe Dry Eye Disease

BACKGROUND: Secretome derived from Wharton’s jelly mesenchymal stem cells (WJ-MSC) has a beneficial effect for ocular surface regeneration; however, the high amount of vascular endothelial growth factor (VEGF) remains a challenge for its application. 
 AIM: The aim of the study was to investigate the effect of resveratrol (RV) (VEGF reducing agent) preconditioned WJ-MSC secretome in Concanavalin A-induced severe dry eye model. 
 METHODS: Pre- and post-experimental study composed of topical instillation of WJ-MSC secretome group, balanced salt solution control group, and normal control group. Tear production, tear break-up time (TBUT), corneal fluorescein dye staining, VEGF level in aqueous tear, goblet cell density, and inflammatory cells in the ocular surface were analyzed. 
 RESULTS: Topical instillation of RV preconditioned WJ-MSC secretome successfully improved tear film production (p = 0.008) and TBUT (p = 0.008), promoted goblet cell restoration (p = 0.023) and reduced corneal fluorescein staining (p = 0.003), while inhibited infiltration of inflammatory cells (p = 0.01) and secretion of VEGF in aqueous tear (p = 0.003). 
 CONCLUSION: Topical instillation of RV preconditioned WJ-MSC secretome has great potential as cell-free based therapy to preserve ocular surface in an experimental model of severe Dry eye disease.

Clinical effects of intrathecal administration of expanded Wharton jelly mesenchymal stromal cells in patients with chronic complete spinal cord injury: a randomized controlled study

Background aimsSpinal cord injury (SCI) represents a devastating condition leading to severe disability related to motor, sensory and autonomic dysfunction. Stem cell transplantation is considered a potential emerging therapy to stimulate neuroplastic and neuroregenerative processes after SCI. In this clinical trial, the authors investigated the safety and clinical recovery effects of intrathecal infusion of expanded Wharton jelly mesenchymal stromal cells (WJ-MSCs) in chronic complete SCI patients. MethodsThe authors designed a randomized, double-blind, crossover, placebo-controlled, phase 1/2a clinical trial (NCT03003364). Participants were 10 patients (7 males, 3 females, age range, 25–47 years) with chronic complete SCI (American Spinal Injury Association A) at dorsal level (T3-11). Patients were randomly assigned to receive a single dose of intrathecal ex vivo-expanded WJ-MSCs (10 × 106 cells) from human umbilical cord or placebo and were then switched to the other arm at 6 months. Clinical evaluation (American Spinal Injury Association impairment scale motor and sensory score, spasticity, neuropathic pain, electrical perception and pain thresholds), lower limb motor evoked potentials (MEPs) and sensory evoked potentials (SEPs), Spinal Cord Independence Measure and World Health Organization Quality of Life Brief Version were assessed at baseline, 1 month, 3 months and 6 months after each intervention. Urodynamic studies and urinary-specific quality of life (Qualiveen questionnaire) as well as anorectal manometry, functional assessment of bowel dysfunction (Rome III diagnostic questionnaire) and severity of fecal incontinence (Wexner score) were conducted at baseline and at 6 months after each intervention. ResultsIntrathecal transplantation of WJ-MSCs was considered safe, with no significant side effects. Following MSC infusion, the authors found significant improvement in pinprick sensation in the dermatomes below the level of injury compared with placebo. Other clinically relevant effects, such as an increase in bladder maximum capacity and compliance and a decrease in bladder neurogenic hyperactivity and external sphincter dyssynergy, were observed only at the individual level. No changes in motor function, spasticity, MEPs, SEPs, bowel function, quality of life or independence measures were observed. ConclusionsIntrathecal transplantation of human umbilical cord-derived WJ-MSCs is a safe intervention. A single intrathecal infusion of WJ-MSCs in patients with chronic complete SCI induced sensory improvement in the segments adjacent to the injury site.

Behavioral Changes in Stem-Cell Potency by HepG2-Exhausted Medium.

Wharton jelly mesenchymal stem cells (WJ-MSCs) are able to differentiate into different cell lineages upon stimulation. This ability is closely related to the perfect balance between the pluripotency-related genes, which control stem-cell proliferation, and genes able to orchestrate the appearance of a specific phenotype. Here we studied the expression of stemness-related genes, epigenetic regulators (DNMT1, SIRT1), miRNAs (miR-145, miR-148, and miR-185) related to stemness, exosomes, the cell-cycle regulators p21 (WAF1/CIP1) and p53, and the senescence-associated genes (p16, p19, and hTERT). Cells were cultured in the presence or absence of the human hepatocarcinoma cell line HepG2-exhausted medium, to evaluate changes in stemness, differentiation capability, and senescence sensibility. Our results showed the overexpression of SIRT1 and reduced levels of p21 mRNA. Moreover, we observed a downregulation of DNMT1, and a simultaneous overexpression of Oct-4 and c-Myc. These findings suggest that WJ-MSCs are more likely to retain a stem phenotype and sometimes to switch to a highly undifferentiable proliferative-like behavior if treated with medium exhausted by human HepG2 cell lines.

Effects of Wharton’s jelly mesenchymal stem cells-derived secretome on colon carcinoma HT-29 cells

Secreted factors (secretome) of Wharton’s jelly mesenchymal stem cells (WJMSCs) have therapeutic impacts. This study was conducted to investigate the impact of WJMSCs-derived secretome (WJMSCs-Se) in apoptosis and the growth of HT-29 cells. HT-29 cells treated with 25 or 50 μg/mL WJMSCs-Se for 24 h. Colony formation and MTT test was used to assess the proliferation and cytotoxicity of the HT-29 cells. Annexin V/PI staining was done for the assessment of apoptosis. The mRNA expression of important apoptosis-related genes was also examined. In the WJMSCs-Se-treated HT-29 cells, colony numbers and viability percentages were significantly reduced in a concentration-dependent manner. Apoptotic and necrotic indexes of WJMSCs-Se-treated HT-29 cells considerably enhanced in comparison to the control. The Caspase-9 and -3 activities were significantly increased in the WJMSCs-Se-exposed HT-29 cells. The mRNA expression of Caspase-9, Caspase-3, and Bax/ Bcl-2 ratio was considerably elevated in the WJMSCs-Se-treated HT-29 cells. Caspase-8 activity and expression of the p53 gene were not affected by the WJMSCs-Se. Taken together, we concluded that WJSCs-Se significantly prevented cell growth and enhanced colon cancer cell death in a concentration-dependent manner mainly through the intrinsic apoptotic pathway.

Static magnetic field-enhanced osteogenic differentiation of human umbilical cord-derived mesenchymal stem cells via matrix vesicle secretion

Background/purpose: Wharton’s jelly mesenchymal stem cells (WJMSCs) are well-known for use in nontissue-specific stem cell therapy. However, they can be a challenge for orthopedic used because they have low osteogenic differentiation ability. Several studies have found that static magnetic fields (SMFs) have positive effects on the osteogenesis of different stem cells. Nevertheless, whether SMFs have similar effects on WJMSC differentiation is unknown. Matrix vesicles are the critical characteristic of osteogenesis. Matrix vesicles serve as the initial site for physiological mineralization. Thus, the purpose of this study was to evaluate the effectiveness of a 0.4-T SMF on osteogenic differentiation and matrix vesicle secretion of WJMSCs. Methods In methodology, WJMSCs were treated with a 0.4-T SMF. The cell viability was tested using the MTT assay. For the osteogenic analysis, the alkaline phosphatase activity assay and alizarin red S staining were performed. The osteogenic-related gene expression of ALP, BMP-2, and Runx2 was examined using real-time polymerase chain reaction. Scanning electron microscopy combined with energy-dispersive X-ray spectroscopy was used to analyze matrix vesicle secretion. Results The cell viability showed no significant difference between the SMF-treated group and the sham-exposed cells. However, the SMF-treated group exhibited significantly more mineralized nodule formation and higher ALP activity than their control counterparts (p < .05). The expressions of osteogenic-related markers, ALP, BMP-2, and Runx2, were also significantly higher in the SMF-treated WJMSCs. The scanning electron microscopy results showed much more matrix vesicle secretion in the SMF-treated cells than in the sham-treated cells. A mineralized sheath was noted in the SMF-treated cells, along with a sporadic accumulation of spherical mineralized deposits on the cell surface. Conclusions The results suggest that 0.4-T SMF treatment enhances the osteogenesis of WJMSCs at the early-to-middle stage of osteogenic differentiation by increasing the matrix vesicle secretion and mineralization.

Comparison of miRNA Cargo within Wharton's Jelly mesenchymal stem cell-derived exosomes manufactured in FBS and hPLT expansion media

Background & Aim Wharton's Jelly Mesenchymal Stem Cell (WJMSCs)-derived exosomes have been studied extensively in animal models of degenerative and chronic diseases. Thus, the development of a large-scale manufacturing procedure to harvest clinical-grade exosomes is required for the translation of exosome to the clinic. We have developed an exosome production technique to manufacture large quantities of exosomes using the quantum bioreactor (Terumo BCT) with both fetal bovine serum (FBS) and human platelet lysate (hPLT) supplemented expansion media. AWe aimed to compare the miRNA cargo of exosomes isolated from FBS-expanded and PLT-expanded WJMSC to determine if growth media is a significant contributor to exosome product preparation. Methods, Results & Conclusion WJMSCs were isolated from Wharton's Jelly and expanded to passage 1. After harvest, 30 × 106 cells were loaded into the quantum bioreactor and expanded for 1 week to achieve maximum confluence. Expansion media was washed and replaced with serum-free aMEM media. Conditioned media was collected every 24 hours for 120 hours and exosomes were precipitated by sequential centrifugation and ultracentrifugation. NanoSight analysis of the final products revealed a production of 15.0 × 1012± 0.32 × 1012 total particles with an average mode size of 111±4.78 nm. Surface protein analysis revealed positive expression of exosome markers CD9, CD63, and CD81. Exosome products were subjected to miRNA isolation, sequencing, and miRNA identification for bioinformatics analysis with the Qiagen IPA software. miRNA analysis was completed using 3 WJMSC donors grown in 20% FBS or 5%PLT supplemented media. Sequencing results identified 157 miRNAs in the PLT group and 154 miRNAs in the FBS group (with a 100-copy minimum expression cut off). FBS and PLT comparison revealed 142 common miRNAs with 15 unique to PLT and 12 unique to FBS. Further RNA target analysis revealed 68 miRNAs that target 1024 mRNA in the PLT group and 64 miRNA that target 1048 mRNA in the FBS group. The two groups share 998 common mRNA targets with 27 unique to PLT and 51 unique to FBS. Canonical pathway analysis of the unique targets reveals the top pathway hits, serotonin receptor and JAK/Stat signaling for PLT and IL-7 and RANK signaling for FBS. We found both expansion medias to be effective for large scale expansion and downstream exosome production. miRNA sequencing revealed small variations in the total number of predicted mRNA targets, suggesting a conserved therapeutic effect between the two groups. Wharton's Jelly Mesenchymal Stem Cell (WJMSCs)-derived exosomes have been studied extensively in animal models of degenerative and chronic diseases. Thus, the development of a large-scale manufacturing procedure to harvest clinical-grade exosomes is required for the translation of exosome to the clinic. We have developed an exosome production technique to manufacture large quantities of exosomes using the quantum bioreactor (Terumo BCT) with both fetal bovine serum (FBS) and human platelet lysate (hPLT) supplemented expansion media. AWe aimed to compare the miRNA cargo of exosomes isolated from FBS-expanded and PLT-expanded WJMSC to determine if growth media is a significant contributor to exosome product preparation. WJMSCs were isolated from Wharton's Jelly and expanded to passage 1. After harvest, 30 × 106 cells were loaded into the quantum bioreactor and expanded for 1 week to achieve maximum confluence. Expansion media was washed and replaced with serum-free aMEM media. Conditioned media was collected every 24 hours for 120 hours and exosomes were precipitated by sequential centrifugation and ultracentrifugation. NanoSight analysis of the final products revealed a production of 15.0 × 1012± 0.32 × 1012 total particles with an average mode size of 111±4.78 nm. Surface protein analysis revealed positive expression of exosome markers CD9, CD63, and CD81. Exosome products were subjected to miRNA isolation, sequencing, and miRNA identification for bioinformatics analysis with the Qiagen IPA software. miRNA analysis was completed using 3 WJMSC donors grown in 20% FBS or 5%PLT supplemented media.

Osteogenic potential of Wharton's jelly mesenchymal stromal cells cultured on a biomimetic scaffold

Background & Aim In regenerative medicine, Wharton's jelly mesenchymal stromal cells (WJ-MSCs) have a growing interest in its properties similar to those of bone marrow cells and the immunomodulatory potential. Tissue engineering combines the design of biomaterials and the potential of cells implanted in the scaffold that interacts with the added growth factors, and its main objective is to return morphology and function by providing a biological mechanically stable niche. This evidence has directed the approach of tissue regeneration towards the use of decellularized native cell matrices. The mature organs and tissues adequately decellularized, can be used as scaffolds, because the immunogenic determinants present in the cells are eliminated, conserving a native MEC, in which its structure, architecture and mechanical properties are preserved, along with the bioavailability of proteins and growth factors necessary for tissue regeneration. We aimed is provide a temporary bone biomimetic scaffold capable of inducing osteogenic differentiation of Wharton's jelly mesenchymal stromal cells. Methods, Results & Conclusion We decellularized, demineralized, and hydrolyzed the collagen protein of the bovine bone. Then we evaluate the structure and decellularization of the scaffolds. We performed scanning electron microscopy and chemical analysis by EDS. Raman Spectroscopy and FTIR techniques determined the chemical composition by identifying functional groups and the fingerprint of type I collagen protein. We aisled WJ-MSCs and characterize surface antigens profiles of WJ-MSCs; we kept the cells in culture for 3,7,14,21 and 28 days to evaluate the biocompatibility of the scaffolds and the cellular response in vitro. On histology sections, the scaffolds showed the conservation of the structure of the ECM and cell removal. We determined that the main component is collagen type I with different degrees of denaturation depending on the concentration of hydrochloric acid (HCl) by Raman and FTIR. We eliminate the mineral phase according to the guidelines established for the FDA for demineralized scaffolds. The scaffolds also showed excellently swelling and mechanical stability. In the in vitro cell culture, the scaffolds have no cytotoxic effect and mineralized matrix deposit after 28 days in culture in the absence of growth factors or inducers of mineralization. Future studies using animal subjects are required to evaluate the degree of mineralization and scaffold response.

A phase i study to evaluate the safety of wharton's jelly mesenchymal stem cells for the treatment of de novo high risk or steroid refractory acute Graft Versus Host Disease (aGVHD)

Background & Aim Allogeneic hematopoietic stem cell transplantation used to manage hematologic malignancies, is frequently complicated by graft versus host disease (GvHD) leading to poor outcomes with increased non-relapse morbidity and mortality. Traditional immunosuppressive therapy has many side effects, thus, necessitating alternative therapies to treat GvHD. Based on their immunomodulation and immunosuppression properties, mesenchymal stem cells (MSCs) are being explored to treat acute GvHD. Currently, there are no cellular products approved in the United States to treat steroid refractory aGvHD. MSCTC-0010, manufactured at the Midwest Stem Cell Therapy Center's GMP facility, is an investigational product consisting of tissue-derived allogeneic, Wharton's Jelly MSCs. We investigated MSCTC-0010 for the treatment of de novo high-risk acute or steroid refractory GvHD in a Phase I, safety clinical trial (IND#17672). The MSCTC-0010 is manufactured by explantation, ex-vivo culture and expansion, and cryopreservation and is tested for identity, potency, sterility, and stability. Methods, Results & Conclusion Method MSCTC-0010 was delivered intravenously twice at a dose of 2 × 106cells/kg in the first cohort and 10 × 106cells/kg in the second cohort on day 0 and 7. GvHD was graded by organ (skin, liver, and gastrointestinal) involvement. The primary endpoint was to evaluate the safety of MSCTC-0010 in patients who experienced a treatment related adverse event. The secondary endpoints include achievement of complete response of aGvHD by day 42 and improvement of GvHD in 1 or more organs involved. Samples were drawn at regular intervals to explore biomarkers that correlate with biological response. Results The study enrolled 10 patients (ages 18-75). Safety of the MSCTC-0010 was demonstrated as there was no infusion related toxicity and no increased incidence of infections. The study showed aGvHD response in 7 of the 10 patients at day 28 (refer to table). A biomarker panel consisting of Reg 3-α and ST-2 was analyzed. Analysis of these two biomarkers showed a reduction from day 0 to day 42. Subjects who received the higher dose of MSCTC-0010 had a statistically significant decline in the Reg-3 α levels on day 7, 14, 21, and 28. The drop in levels correlated with the clinical response. ST-2 levels showed a significant decrease on day 28 and 42. Conclusion This Phase 1 trial with MSCTC-0010 successfully met its primary endpoint of safety. Treatment with MSCTC-0010 shows promise in patients with high risk aGvHD.

Intranasal administration of conditioned medium derived from mesenchymal stem cells-differentiated oligodendrocytes ameliorates experimental autoimmune encephalomyelitis

In multiple sclerosis, myelin sheaths around the axons are degenerated due to uncontrolled inflammation in the central nervous system. Oligodendrocytes (OLs) are myelin-forming cells that secrete trophic factors necessary for myelin protection. Beneficial features of conditioned medium (CM) derived from different stem cells are nowadays under investigation in treating neurodegenerative diseases. Here, we used the differentiation capacity of Wharton's jelly mesenchymal stem cells (WJMSCs) to obtain OLs. Then, the study aimed to evaluate the status of inflammation and myelination in male experimental autoimmune encephalomyelitis (EAE) mice after intranasal administration of CM derived from OLs (OL-CM). Inflammation was studied by evaluating gliosis, inflammatory cell infiltration and expression of inflammation indicators including NLRP3 inflammasome, interleukin-1β, interleukin-18, glial fibrillary acidic protein, and ionized calcium binding adaptor molecule 1. Remyelination was studied by luxol fast blue staining and evaluating the expression of myelin indicators including myelin basic protein and oligodendrocyte transcription factor. In addition, we followed the trend of body weight and functional recovery during the 28-day study. ELISA assay revealed that OL-CM contained brain-derived neurotrophic factor, glial cell-derived neurotrophic factor, and ciliary neurotrophic factor. Data showed that OL-CM moderated inflammation, augmented remyelination, and gained normal body weight. Notably, these anti-inflammatory and regenerative effects of OL-CM improved neurological functions in EAE mice. In conclusion, the current study offered a new choice for treating multiple sclerosis using noninvasive intranasal administration of CM harvested from easily achievable WJMSCs-differentiated OLs.

Adjustment of Cell Adhesion on Polyurethane Structures via Control of the Hard/Soft Segment Ratio

AbstractCreating substrates with a similar composition that can either prevent or promote cell adhesion is still a challenging feat. Here, it is shown that a strikingly simple method of tuning the amount of hard segments or isocyanate index (NCOind) of a polyurethane (PU) film allows to modulate cell adhesiveness. PU films are synthesized with NCOind of 75, 100, 200, 300 and 400 corresponding to ratios of isocyanate to hydroxyl functions of 0.75, 1, 2, 3, 4, respectively. The adhesive capacity of NIH 3T3 fibroblasts (3T3) and Wharton's jelly mesenchymal stem cells (WJMSCs) are dependent on the NCOind. For NCOind below 300, no cell adhesion can be observed regardless of the cell type, whereas for NCOind of 300 and 400 cells adhere to the PU surface. WAXS and small angle X‐ray scattering (SAXS) studies reveal that variations of NCOind allows to modulate the phase separation in PU films. Porod's law shows that for NCOind of 300 and 400, the hard–soft segment interface is sharp. Conversely, samples with smaller NCOind present diffuse interfaces. Hence, the morphology of the interface between hard and soft domains appears to be a critical feature that correlates with the adhesion capacity of cells.

Xenogeneic transplantation of human WJ-MSCs rescues mice from acute radiation syndrome via Nrf-2-dependent regeneration of damaged tissues.

There is an unmet medical need for radiation countermeasures that can be deployed for treatment of exposed individuals during ionizing radiation (IR) accidents or terrorism. Wharton's jelly mesenchymal stem cells (WJ-MSCs) from human umbilical cord have been shown to avoid allorecognition and induce a tissue-regenerating microenvironment, which makes them an attractive candidate for mitigating IR injury. We found that WJ-MSCs protected mice from a lethal dose of IR even when transplanted up to 24hours after irradiation, and a combination of WJ-MSCs and antibiotic (tetracycline) could further expand the window of protection offered by WJ-MSCs. This combinatorial approach mitigated IR-induced damage to the hematopoietic and gastrointestinal system. WJ-MSCs increased the serum concentration of the cytoprotective cytokines granulocyte colony-stimulating factor (G-CSF) and IL-6 in mice. Knockdown of G-CSF and IL-6 in WJ-MSCs before injection to lethally irradiated mice or transplantation of WJ-MSCs to lethally irradiated Nrf-2 knockout mice significantly nullified the therapeutic protective efficacy. Hence, WJ-MSCs could be a potential cell-based therapy for individuals accidentally exposed to radiation.

Potential of ovine Wharton jelly derived mesenchymal stem cells to transdifferentiate into neuronal phenotype for application in neuroregenerative therapy

Introduction: The transdifferentiation potential of mesenchymal stem cells (MSCs) is not limited to mesodermal derivatives but also to other cell types such as neuronal cells under appropriate cell culture conditions.Materials and methods: The present study characterizes the differentiation of Wharton’s jelly (WJ) derived MSCs using neuronal conditioned medium (NCM) collected from cultured foetal brain cells.Results: After induction with NCM to neuronal stem cells (NSC), the WJ MSCs showed profound morphological changes showing multiple neurites extending from the cell body containing reminiscent of Nissl substance and single long axon-like processes. In RT PCR and immunocytochemistry, the induced neuronal cells showed a strong positive expression of neuronal markers Nestin, β III tubulin and GFAP indicated that, the cells were reactive to NCM for differentiation. A significant (p < 0.01) increase in the level of secretome BDNF was observed in NCM suggests that the BDNF could play a key role in the transdifferentiation of WJMSCs to NSCs.Conclusion: These results support the potential of ovine MSCs isolated from umbilical cord WJ of abattoir derived foetuses to differentiate into neuronal stem cells and also provide a valuable experimental data for NSC transplant research in veterinary medicine.

Wharton’s Jelly MSCs: Potential Weapon to Sharpen for Our Battle against DM

Diabetes mellitus (DM) is a devastating metabolic disease. Stem cell therapy provides great hope to all diabetic patients. Umbilical cord (UC) Wharton's jelly mesenchymal stem cells (WJ-MSCs) specifically provides a potential cell therapy for DM. In this article, we discuss major advantages of WJ-MSCs and challenges facing their clinical utility in DM.

Wharton's Jelly Mesenchymal Stem Cell: Various Protocols for Isolation and Differentiation of Hepatocyte-Like Cells; Narrative Review.

There are several differentiation methods for mesenchymal stem cells (MSCs) into hepatocyte-like cell. Investigators reported various hepatic differentiation protocols such as modifying culturing conditions or using various growth factors/cytokines. In this literature review, we compared different MSCs extraction and isolation protocols from Wharton’s jelly (WJ) and explored various MSCs differentiation methods. Various protocols have been recommended for MSCs isolated from WJ, such as enzymatic, enzymatic-explant, and explant methods. In the explant method, valuable time is wasted, but the cost and biological contaminations are reduced and the number of isolated cells is high. However, other features, such as immune phenotype and multiline-age differentiation capacity, do not differ from other methods. There are also several differentiation methods for hepatocyte-like cell including the induction of MSC by cytokines and growth factors, and the differentiation of MSC in 2- and 3-dimensional matrix (2D and 3D). Among several cytokines, hepatocyte growth factor (HGF) and fibroblast growth factor (FGF) are essential. In the early stage of the differentiation, 2D culture is useful, and in the development stage, 3D culture system with HGF and FGF cytokines are more effective in the process of differentiation. Some studies have used 3D culture system in biocompatible scaffolds, such as alginate, collagen, gelatin, and peptide-Gly-Leu-amide (PGLA). In conclusion, Wharton’s jelly-Mesenchymal stem cells (WJ-MSCs) can be considered as an appropriate source for hepatocyte differentiation. Moreover, we introduced the explant method as the most effective protocol. This review attempted to highlight factors in hepatocyte differentiation, but the most effective protocol is not still unknown.

Sulfur and nitrogen containing plasma polymers reduces bacterial attachment and growth.

Role of sulfur (S) and nitrogen (N) groups in promoting cell adhesion or commonly known as biocompatibility, is well established, but their role in reducing bacterial attachment and growth is less explored or not well-understood. Natural sulfur-based compounds, i.e. sulfide, sulfoxide and sulfinic groups, have shown to inhibit bacterial adhesion and biofilm formation. Hence, we mimicked these surfaces by plasma polymerizing thiophene (ppT) and air-plasma treating this ppT to achieve coatings with S of similar oxidation states as natural compounds (ppT-air). In addition, the effects of these N and S groups from ppT-air were also compared with the biocompatible amine-amide from n-heptylamine plasma polymer. Crystal violet assay and live and dead fluorescence staining of E. coli and S. aureus showed that all the N and S coated surfaces generated, including ppHA, ppT and ppT-air, produced similarly potent, growth reduction of both bacteria by approximately 65% at 72 h compared to untreated glass control. The ability of osteogenic differentiation in Wharton's jelly mesenchymal stem cells (WJ-MSCs) were also used to test the cell biocompatibility of these surfaces. Alkaline phosphatase assay and scanning electron microscopy imaging of these WJ-MSCs growths indicated that ppHA, and ppT-air were cell-friendly surfaces, with ppHA showing the highest osteogenic activity. In summary, the N and S containing surfaces could reduce bacteria growth while promoting mammalian cell growth, thus serve as potential candidate surfaces to be explored further for biomaterial applications.

The suspension culture method to induce Wharton’s jelly mesenchymal stem cells (WJMSCs) into islets-like clusters and in vivo transplantation to control blood glucose of diabetic nude mice

The suspension culture method to induce Wharton’s jelly mesenchymal stem cells (WJMSCs) into islets-like clusters and in vivo transplantation to control blood glucose of diabetic nude mice