Cord Tissue-derived Mesenchymal Stromal Cells Research Articles

Repeated intravenous doses of human umbilical cord-derived mesenchymal stromal cells for bronchopulmonary dysplasia: results of a phase 1 clinical trial with 2-year follow-up

BackgroundCurrently, there is a lack of effective treatments or preventive strategies for bronchopulmonary dysplasia (BPD). Pre-clinical studies with mesenchymal stromal cells (MSCs) have yielded encouraging results. The safety of administering repeated intravenous doses of umbilical cord tissue-derived mesenchymal stromal cells (UC-MSCs) has not yet been tested in extremely-low-gestational-age newborns (ELGANs). Aimsto test the safety and feasibility of administering three sequential intravenous doses of UC-MSCs every 7 days to ELGANs at risk of developing BPD. Methods: In this phase 1 clinical trial, we recruited ELGANs (birth weight ≤1250 g and ≤28 weeks in gestational age [GA]) who were on invasive mechanical ventilation (IMV) with FiO2 ≥ 0.3 at postnatal days 7–14. Three doses of 5 × 106/kg of UC-MSCs were intravenously administered at weekly intervals. Adverse effects and prematurity-related morbidities were recorded. ResultsFrom April 2019 to July 2020, 10 patients were recruited with a mean GA of 25.2 ± 0.8 weeks and a mean birth weight of 659.8 ± 153.8 g. All patients received three intravenous UC-MSC doses. The first dose was administered at a mean of 16.6 ± 2.9 postnatal days. All patients were diagnosed with BPD. All patients were discharged from the hospital. No deaths or any serious adverse events related to the infusion of UC-MSCs were observed during administration, hospital stays or at 2-year follow-up. ConclusionsThe administration of repeated intravenous infusion of UC-MSCs in ELGANs at a high risk of developing BPD was feasible and safe in the short- and mid-term follow-up.

Cell-based versus corticosteroid injections for knee pain in osteoarthritis: a randomized phase 3 trial

Various types of cellular injection have become a popular and costly treatment option for patients with knee osteoarthritis despite a paucity of literature establishing relative efficacy to each other or corticosteroid injections. Here we aimed to identify the safety and efficacy of cell injections from autologous bone marrow aspirate concentrate, autologous adipose stromal vascular fraction and allogeneic human umbilical cord tissue-derived mesenchymal stromal cells, in comparison to corticosteroid injection (CSI). The study was a phase 2/3, four-arm parallel, multicenter, single-blind, randomized, controlled clinical trial with 480 patients with a diagnosis of knee osteoarthritis (Kellgren–Lawrence II–IV). Participants were randomized to the three different arms with a 3:1 distribution. Arm 1: autologous bone marrow aspirate concentrate (n = 120), CSI (n = 40); arm 2: umbilical cord tissue-derived mesenchymal stromal cells (n = 120), CSI (n = 40); arm 3: stromal vascular fraction (n = 120), CSI (n = 40). The co-primary endpoints were the visual analog scale pain score and Knee injury and Osteoarthritis Outcome Score pain score at 12 months versus baseline. Analyses of our primary endpoints, with 440 patients, revealed that at 1 year post injection, none of the three orthobiologic injections was superior to another, or to the CSI control. In addition, none of the four groups showed a significant change in magnetic resonance imaging osteoarthritis score compared to baseline. No procedure-related serious adverse events were reported during the study period. In summary, this study shows that at 1 year post injection, there was no superior orthobiologic as compared to CSI for knee osteoarthritis. ClinicalTrials.gov Identifier: NCT03818737

HLA-G and CD152 Expression Levels Encourage the Use of Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells as an Alternative for Immunosuppressive Therapy.

Mesenchymal stromal cells (MSCs) have been used in immunosuppressive therapy due to their therapeutic effects, with the HLA-G molecule seeming to play a fundamental role. This work evaluated alternative MSC sources to bone marrow (BM), namely, umbilical cord tissue (UC), adipose tissue (AD) and dental pulp tissue (DP), and the influence of interferon-γ (IFN-γ) and hypoxia on the cultivation of these cells for use in immunosuppression therapies. Expression of costimulatory markers CD40, CD80 and CD86 and immunosuppressive molecules CD152 and HLA-G was analyzed. Lymphocyte inhibition assays were also performed. Sequencing of the HLA-G gene from exons 1 to 5 was performed using next-generation sequencing to determine the presence of alleles. UC-derived MSCs (UCMSCs) expressed higher CD152 and HLA-G1 under standard cultivation. UCMSCs and DP-derived MSCs (DPSCs) secreted similar levels of HLA-G5. All MSC sources inhibited the proliferation of peripheral blood mononuclear cells (PBMCs); growth under regular versus hypoxic conditions resulted in similar levels of inhibition. When IFN-γ was added, PBMC growth was inhibited to a lesser extent by UCMSCs. The HLA-G*01:04:01:01 allele appears to generate a more efficient MSC response in inhibiting lymphocyte proliferation. However, the strength of this conclusion was limited by the small sample size. UCMSCs are an excellent alternative to BM in immunosuppressive therapy: they express high concentrations of inhibitory molecules and can be cultivated without stimuli, which minimizes cost.

A critical appraisal of humanized alternatives to fetal bovine serum for clinical applications of umbilical cord derived mesenchymal stromal cells.

The study is aimed to verify the possibility of using humanized alternatives to fetal bovine serum (FBS) such as umbilical cord blood plasma (CBP) and AB+ plasma to support the long-term growth of mesenchymal stromal cells (MSCs) derived from the umbilical cord. We hypothesized that umbilical CBP would be a potential substitute to FBS, especially for small scale autologous clinical transplantations. The MSCs were cultured for six consecutive passages to evaluate xeno-free media's ability to support long-term growth. Cell proliferation rates, colony-forming-unit (CFU) efficiency and population doublings of expanded MSCs, were investigated. Ex vivo expanded MSCs were further characterized using flow cytometry and quantitative PCR. The impact of cryopreservation and composition of cryomedium on phenotype, viability of MSC was also assessed. Our results on cell proliferation, colony-forming unit efficiency suggested that the expansion of the cells was successfully carried out in media supplemented with humanized alternatives. MSCs showed lower CFU counts in FBS (~ 25) than humanized alternatives (~ 35). The gene expression analysis revealed that transcripts showed significant differential expression by two to three folds in the FBS group compared with MSCs grown in medium with humanized alternatives (p < 0.05). In addition, MSCs grown in a medium with FBS had more osteogenic activity, a signatureof unwanted differentiation. The majority of ex vivo expanded MSCs at early and late passages expressed CD44+, CD73+, CD105+, CD90+, and CD166+ in all the experimental groups tested (~ 90%). In contrast to the other MSC surface markers, expression levels of STRO-1+ (~ 21-10%) and TNAP+ (~ 29-11%) decreased with the increase in passage number for MSCs cultured in a FBS-supplemented medium (p < 0.05). Our results established that CBP supported culture of umbilical cord tissue-derived MSCs and is a safer Xeno free replacement to FBS. The use of CBP also enables the storage of umbilical cord tissue derived MSCs in patient-specific conditions to minimize adverse events if cells are delivered directly to the patient.

In vitro evaluation of umbilical cord tissue-derived mesenchymal stromal cells to alleviate occupational lung disease

In vitro evaluation of umbilical cord tissue-derived mesenchymal stromal cells to alleviate occupational lung disease

USE OF TOCILIZUMAB AND MESENCHYMAL STROMAL CELLS IN THE TREATMENT OF SEVERE COVID-19 - A CASE REPORT

BackgroundCurrently, there is no specific treatment for coronavirus disease, and some drugs and cell-based therapy have been tested as alternatives. This work aims to evaluate the effects of the combined use of humanized recombinant monoclonal antibody capable of binding the IL-6 receptor (Tocilizumab), and umbilical cord tissue-derived mesenchymal stromal cells (UCT-MSC) in the treatment of a patient with severe COVID-19 admitted to the intensive care unit (IUC) and submitted to mechanical ventilation.MethodsThis study is part of a project approved by the National Research Ethics Commission (CONEP); CAAE: 30833820.8.0000.0020. The patient had a diagnostic criterion for the severe acute respiratory syndrome resulting from infection with SARS-CoV-2 and received two 400 mg doses of tocilizumab, three infusions of 500,000 CTM / kg plus full anticoagulation. TCU-MSC were obtained from healthy donors. The following parameters were evaluated in the pre-infusion of cells (D1), on the day following each infusion (D2, D4, and D6), on the 14th and 60th day after the first infusion (D14 and D60): viral load, immune response (Regulatory T lymphocytes), C-reactive protein level in plasma, oxygen saturation, respiratory rate, total lymphocyte count and subpopulations (platelets, inflammatory cells, and reticulocytes), TGO / TGP, increased prothrombin time, D-dimer, creatinine, troponin.ResultsThe relative viral quantification decreased gradually from 1 (D1) to 0.06 (D6) RdRP / RNApol, undetectable in D14. An increase in the absolute number of total lymphocytes / µL has also been seen to have progressively increased from 281 (D1) to 954.9 (D6) and since then decreased to 641.6 in D60 in the same way as T lymphocytes 148.6 (D1) 642.6 (D6) 607.4 (D14) 485.7 (D60), CD4 T lymphocytes, 102 (D1) 481.2 (D6) 459.5 (D14) 358 (D60) and Treg lymphocytes 10.8 (D1) 34 (D6) 29.8 (D14), 25.9 (D60). Plasmablasts, in contrast, decreased from 52 (D1) to 4.5 (D6) to almost undetectable in D60 (0.2). Laboratory tests outside the reference values decreased during the follow-up from D1 to D14 were within the normal parameters at D60. The patient was extubated uneventfully on D6, discharged from the ICU on D10, and the hospital on D14.Conclusionthe combined use of tocilizumab and MSC is safe, without adverse effects, and the results of this case report prove to be a promising alternative in the treatment of patients with severe acute respiratory syndrome due to SARS-CoV-2.

Cord Blood Connect: The International Congress for Cord Blood and Perinatal Tissue Research 2020.

The proceedings contain 15 papers The topics discussed include: targeting neuroinflammation with human umbilical cord tissue-derived mesenchymal stromal cells;multiple doses of umbilical cord blood cells improve long-term perinatal brain injury;COVID-19 and its impact on collection and processing of cord blood units at the Cleveland Cord Blood Center;hydrocortisone-treated DUOC-01, a cord blood-derived cell therapy product, ameliorates experimental autoimmune encephalomyelitis;measuring microglial suppression with multiple assays;impact of donor age on outcomes in hematopoietic stem cell transplantation;single UM171-expanded cord blood transplants support robust T-cell reconstitution with low rates of severe infections;autologous umbilical cord blood-derived cell administration in extreme preterm infants: CORD-SAFE study;and umbilical cord blood infusion reactions

Comparison of umbilical cord tissue-derived mesenchymal stromal cells isolated from cryopreserved material and extracted by explantation and digestion methods utilizing a split manufacturing model

Background aimsUmbilical cord (UC) tissue is recognized as an advantageous source of mesenchymal stromal cells (MSCs), whose therapeutic properties are being actively evaluated in pre-clinical and clinical trials. In recognition of its potential value, storage of UC tissue or cells from UC tissue in newborn stem cell banks is now commonplace; however, strategies for isolating UC-derived MSCs (UCMSCs) from UC tissue have not been standardized. The majority of newborn stem cell banks take one of two approaches to cord tissue processing and cryopreservation: enzymatic digestion of the fresh tissue with cryopreservation of the subsequent cell suspension or cryopreservation of the tissue as a composite whole with later, post-thaw isolation of cells by explantation. Evaluation of UCMSCs derived by these two principal preparation and cryopreservation strategies is important to understanding whether the methods currently employed by newborn stem cell banks retain the desirable clinical attributes of UC cells. MethodsUCMSCs were isolated from 10 UC tissue samples by both explantation and enzymatic digestion methods to allow for comparison of cells from the same donor. Cell isolates from both methods were compared pre- and post-cryopreservation as well as after serial passaging. Cell viability, morphology, growth kinetics, immunophenotype, cytokine secretion and differentiation capacity were evaluated. ResultsUCMSCs could be derived from fresh UC tissue by both explantation and digestion methods and from thawed UC tissue by explantation. Initial cell populations isolated by digestion were heterogeneous and took longer to enrich for UCMSCs in culture than populations obtained by explantation. However, once isolated and enriched, UCMSCs obtained by either method showed no significant difference in viability, morphology, rate of proliferation, surface marker expression, levels of cytokine secretion or differentiation capacity. ConclusionsDerivation of UCMSCs by explantation after thawing UC cryopreserved as a composite tissue may be favorable in terms of initial purity and number of cells achievable by a specific passage. However, we observed no evidence of functional difference between UCMSCs derived by explanation or digestion, suggesting that cells isolated from cryopreserved material obtained by either method maintain their therapeutic properties.

Phase I Trial of Allogeneic Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells in Neonates with Hypoxic-Ischemic Encephalopathy

Background & Aim Background In neonatal animal models of hypoxic-ischemic brain injury administration of mesenchymal stromal cells (MSCs) improves anatomic and functional outcomes. A prior study suggests potential benefit of autologous cord blood-derived cells for infants with hypoxic-ischemic encephalopathy (HIE) but cord blood collection in this population is challenging. A safe, effective off-the-shelf allogeneic cell product could be a useful alternative. Our aim was to conduct a phase I open-label study of umbilical cord tissue-derived MSCs in infants treated with hypothermia for HIE. Methods, Results & Conclusion Methods Infants >35 weeks gestation treated with hypothermia for HIE based on NICHD hypothermia trial criteria were eligible. Goal enrollment was 6 infants. Allogeneic umbilical cord tissue-derived mesenchymal stromal cells (hCT-MSC) were manufactured in the Robertson GMP Cell Manufacturing Laboratory from a single cord tissue donated to the Carolinas Cord Blood Bank. Cells were expanded to passage 2 and cryopreserved. A representative cryobag was quality-tested for cell count, viability, phenotype, functional assays, gram stain and sterility. For administration cells were thawed and diluted 1:1 and delivered in syringes containing plasmalyte-A, 5% HSA, and residual DMSO. Post thaw samples were tested for cell count, viability (required ≥70%) and sterility. The first 3 infants (cohort 1) received 2 × 106 cells/kg i.v. in the first 48 postnatal hours, the second 3 (cohort 2) received a 2nd dose at 2 months. Primary safety outcomes were occurrence of infusion reactions or infections within 2 weeks of infusion. Results Of 18 eligible infants, 6 were enrolled (Table 1). All cultures obtained from subjects were negative, and no subject had an infusion reaction. Subject #5, had a positive panel reactive antibody (PRA) screen at 6 months. Broth culture from thawed hCT-MSC for subject # 1 grew coagulase-negative staphylococci. The infant remained asymptomatic after re-warming and the infant's blood cultures were sterile. The broth culture was assessed as contaminant via handling post-thaw rather than cell product contamination. There was one protocol deviation (One infant received a study dose at 71 hours. (Table 2)). Conclusions Our results suggest that hCT-MSC, an umbilical cord tissue-derived MSC product, can be prepared and infused safely in infants treated with hypothermia for moderate - severe HIE. Further safety and efficacy studies are warranted.

Targeting Neuroinflammation with Human Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells

Targeting Neuroinflammation with Human Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells

Comparative Evaluation of Two Different Xeno-Free Media Supplements in Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells In Vitro Culture

Comparative Evaluation of Two Different Xeno-Free Media Supplements in Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells In Vitro Culture

Meeting the demands of msc production for clinical studies

Background & Aim The Marcus Center for Cellular Cures (MC3) at Duke University recently completed a Phase I clinical trial (NCT03099239) using human cord tissue-derived mesenchymal stromal cells (hCT-MSCs) for the treatment of autism spectrum disorder. For this trial, 12 patients ages 2 – 11 years were sequentially dosed with 1, 2, or 3 doses of 2 million MSC/kg post thaw administered intravenously every 2 months. A manufacturing process was developed for our GMP facility to produce the required doses in a flexible and reproducible manner. Methods, Results & Conclusion For the trial, 3 lots of hCT-MSCs were manufactured from cord tissue obtained from 3 term healthy male babies delivered after elective routine C-section. After transporting the cord to the Robertson GMP Laboratory, the tissue was cut and digested in a tissue dissociator. The resulting cell suspension was plated in tissue culture flasks for P0 culture in Prime-XV MSC Expansion XSFM culture medium (Irvine Scientific) supplemented with 1% platelet lysate (PL). After harvest, P0 cells were cryopreserved, with a subset of cells expanded for P1 in XSFM without PL, and with a larger cryopreservation and expansion process for P2. For both P1 and P2, cells were seeded at 1000/cm2. Cells at the end of P2 were frozen in multi-compartment cryo-bags at the appropriate concentration for final dose administration. Prior to release to the patient, each lot of P2 hCT-MSC was characterized by flow cytometry for typical MSC markers, tri-lineage differentiation, and the ability to suppress T-cell proliferation. Manufacturing process data for each donor are presented in Table 1. For 2 donors, additional P2 cells were produced from P1 vials during the course of the trial (e.g., “remanufactured” lots) when the original batch was not enough to meet demand. We calculated that after P0 the cells underwent 10.5 doublings on average after the end of P2, with a mean doubling time in P1 – P2 of 31.2 hours. The final thawed and prepared P2 cell product had a mean viability of 75% prior to administration to the patient. Preparations are underway for a Phase II clinical trial and the subsequent need for a larger scale and more efficient manufacturing process. We are developing a streamlined process using closed systems for cell expansion, concentration, and cryopreservation of the MSC product that is semi-automated and will minimize the use of clean room facilities. Comparability of the MSC product using the new process will be confirmed prior to use in the Phase II trial.

Targeting Neuroinflammation with Human Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells

Targeting Neuroinflammation with Human Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells

307 - Targeting neuroinflammation with human umbilical cord tissue-derived mesenchymal stromal cells

307 - Targeting neuroinflammation with human umbilical cord tissue-derived mesenchymal stromal cells

Human Umbilical Cord Mesenchymal Stromal Cells Support Viability of Umbilical Cord Blood Hematopoietic Stem Cells but not the "Stemness" of Their Progeny in Co-Culture.

Cell-cell interactions and the ability of mesenchymal stromal cells to support the expansion of hematopoietic progenitor cells were studied in co-culture of human umbilical cord tissue-derived mesenchymal stromal cells and nucleated umbilical cord blood cells. It was found that hematopoietic stem cells from the umbilical cord blood are capable to adhere to mesenchymal stromal cells and proliferate during 3-4 weeks in co-culture. However, despite the formation of hematopoietic foci and accumulation of CD34+ and CD133+ cells in the adherent cell fraction, the ability of newly generated blood cells to form colonies in semi-solid culture medium was appreciably reduced. These findings suggest that human umbilical cord tissue-derived mesenchymal stromal cells display a weak capability to support the "stemness" of hematopoietic stem cell progeny despite long-term maintenance of their viability and proliferation.

Therapeutic angiogenesis induced by human umbilical cord tissue-derived mesenchymal stromal cells in a murine model of hindlimb ischemia.

BackgroundMesenchymal stem cells derived from human umbilical cord tissue, termed UCX®, have the potential to promote a full range of events leading to tissue regeneration and homeostasis. The main goal of this work was to investigate UCX® action in experimentally induced hindlimb ischemia (HLI).MethodsUCX®, obtained by using a proprietary technology developed by ECBio (Amadora, Portugal), were delivered via intramuscular injection to C57BL/6 females after unilateral HLI induction. Perfusion recovery, capillary and collateral density increase were evaluated by laser doppler, CD31 immunohistochemistry and diaphonisation, respectively. The activation state of endothelial cells (ECs) was analysed after EC isolation by laser capture microdissection microscopy followed by RNA extraction, cDNA synthesis and quantitative RT-PCR analysis. The UCX®-conditioned medium was analysed on Gallios flow cytometer. The capacity of UCX® in promoting tubulogenesis and EC migration was assessed by matrigel tubule formation and wound-healing assay, respectively.ResultsWe demonstrated that UCX® enhance angiogenesis in vitro via a paracrine effect. Importantly, after HLI induction, UCX® improve blood perfusion by stimulating angiogenesis and arteriogenesis. This is achieved through a new mechanism in which durable and simultaneous upregulation of transforming growth factor β2, angiopoietin 2, fibroblast growth factor 2, and hepatocyte growth factor, in endothelial cells is induced by UCX®.ConclusionsIn conclusion, our data demonstrate that UCX® improve the angiogenic potency of endothelial cells in the murine ischemic limb suggesting the potential of UCX® as a new therapeutic tool for critical limb ischemia.

Three-dimensional spheroid cell culture of umbilical cord tissue-derived mesenchymal stromal cells leads to enhanced paracrine induction of wound healing.

IntroductionThe secretion of trophic factors by mesenchymal stromal cells has gained increased interest given the benefits it may bring to the treatment of a variety of traumatic injuries such as skin wounds. Herein, we report on a three-dimensional culture-based method to improve the paracrine activity of a specific population of umbilical cord tissue-derived mesenchymal stromal cells (UCX®) towards the application of conditioned medium for the treatment of cutaneous wounds.MethodsA UCX® three-dimensional culture model was developed and characterized with respect to spheroid formation, cell phenotype and cell viability. The secretion by UCX® spheroids of extracellular matrix proteins and trophic factors involved in the wound-healing process was analysed. The skin regenerative potential of UCX® three-dimensional culture-derived conditioned medium (CM3D) was also assessed in vitro and in vivo against UCX® two-dimensional culture-derived conditioned medium (CM2D) using scratch and tubulogenesis assays and a rat wound splinting model, respectively.ResultsUCX® spheroids kept in our three-dimensional system remained viable and multipotent and secreted considerable amounts of vascular endothelial growth factor A, which was undetected in two-dimensional cultures, and higher amounts of matrix metalloproteinase-2, matrix metalloproteinase-9, hepatocyte growth factor, transforming growth factor β1, granulocyte-colony stimulating factor, fibroblast growth factor 2 and interleukin-6, when compared to CM2D. Furthermore, CM3D significantly enhanced elastin production and migration of keratinocytes and fibroblasts in vitro. In turn, tubulogenesis assays revealed increased capillary maturation in the presence of CM3D, as seen by a significant increase in capillary thickness and length when compared to CM2D, and increased branching points and capillary number when compared to basal medium. Finally, CM3D-treated wounds presented signs of faster and better resolution when compared to untreated and CM2D-treated wounds in vivo. Although CM2D proved to be beneficial, CM3D-treated wounds revealed a completely regenerated tissue by day 14 after excisions, with a more mature vascular system already showing glands and hair follicles.ConclusionsThis work unravels an important alternative to the use of cells in the final formulation of advanced therapy medicinal products by providing a proof of concept that a reproducible system for the production of UCX®-conditioned medium can be used to prime a secretome for eventual clinical applications.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-015-0082-5) contains supplementary material, which is available to authorized users.

What Makes Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells Superior Immunomodulators When Compared to Bone Marrow Derived Mesenchymal Stromal Cells?

MSCs derived from the umbilical cord tissue, termed UCX, were investigated for their immunomodulatory properties and compared to bone marrow-derived MSCs (BM-MSCs), the gold-standard in immunotherapy. Immunogenicity and immunosuppression were assessed by mixed lymphocyte reactions, suppression of lymphocyte proliferation and induction of regulatory T cells. Results showed that UCX were less immunogenic and showed higher immunosuppression activity than BM-MSCs. Further, UCX did not need prior activation or priming to exert their immunomodulatory effects. This was further corroborated in vivo in a model of acute inflammation. To elucidate the potency differences observed between UCX and BM-MSCs, gene expression related to immune modulation was analysed in both cell types. Several gene expression profile differences were found between UCX and BM-MSCs, namely decreased expression of HLA-DRA, HO-1, IGFBP1, 4 and 6, ILR1, IL6R and PTGES and increased expression of CD200, CD273, CD274, IL1B, IL-8, LIF and TGFB2. The latter were confirmed at the protein expression level. Overall, these results show that UCX seem to be naturally more potent immunosuppressors and less immunogenic than BM-MSCs. We propose that these differences may be due to increased levels of immunomodulatory surface proteins such as CD200, CD273, CD274 and cytokines such as IL1β, IL-8, LIF and TGFβ2.

Human umbilical cord tissue-derived mesenchymal stromal cells attenuate remodeling after myocardial infarction by proangiogenic, antiapoptotic, and endogenous cell-activation mechanisms

IntroductionAmong the plethora of cells under investigation to restore a functional myocardium, mesenchymal stromal cells (MSCs) have been granted considerable interest. However, whereas the beneficial effects of bone marrow MSCs (BM-MSCs) in the context of the diseased heart are widely reported, data are still scarce on MSCs from the umbilical cord matrix (UCM-MSCs). Herein we report on the effect of UCM-MSC transplantation to the infarcted murine heart, seconded by the dissection of the molecular mechanisms at play.MethodsHuman umbilical cord tissue-derived MSCs (UCX®), obtained by using a proprietary technology developed by ECBio, were delivered via intramyocardial injection to C57BL/6 females subjected to permanent ligation of the left descending coronary artery. Moreover, medium produced by cultured UCX® preconditioned under normoxia (CM) or hypoxia (CMH) was collected for subsequent in vitro assays.ResultsEvaluation of the effects upon intramyocardial transplantation shows that UCX® preserved cardiac function and attenuated cardiac remodeling subsequent to myocardial infarction (MI). UCX® further led to increased capillary density and decreased apoptosis in the injured tissue. In vitro, UCX®-conditioned medium displayed (a) proangiogenic activity by promoting the formation of capillary-like structures by human umbilical vein endothelial cells (HUVECs), and (b) antiapoptotic activity in HL-1 cardiomyocytes subjected to hypoxia. Moreover, in adult murine cardiac Sca-1+ progenitor cells (CPCs), conditioned medium enhanced mitogenic activity while activating a gene program characteristic of cardiomyogenic differentiation.ConclusionsUCX® preserve cardiac function after intramyocardial transplantation in a MI murine model. The cardioprotective effects of UCX® were attributed to paracrine mechanisms that appear to enhance angiogenesis, limit the extent of the apoptosis, augment proliferation, and activate a pool of resident CPCs. Overall, these results suggest that UCX® should be considered an alternative cell source when designing new therapeutic approaches to treat MI.

The role of human umbilical cord tissue-derived mesenchymal stromal cells (UCX®) in the treatment of inflammatory arthritis

BackgroundECBio has developed proprietary technology to consistently isolate, expand and cryopreserve a well-characterized population of stromal cells from human umbilical cord tissue (UCX® cells). The technology has recently been optimized in order to become compliant with Advanced Medicine Therapeutic Products. In this work we report the immunosuppressive capacity of UCX® cells for treating induced autoimmune inflammatory arthritis.MethodsUCX® cells were isolated using a proprietary method (PCT/IB2008/054067) that yields a well-defined number of cells using a precise proportion between tissue digestion enzyme activity units, tissue mass, digestion solution volume and void volume. The procedure includes three recovery steps to avoid non-conformities related to cell recovery. UCX® surface markers were characterized by flow cytometry and UCX® capacity to expand in vitro and to differentiate into adipocyte, chondrocyte and osteoblast-like cells was evaluated. Mixed Lymphocyte Reaction (MLR) assays were performed to evaluate the effect of UCX® cells on T-cell activation and Treg conversion assays were also performed in vitro. Furthermore, UCX® cells were administered in vivo in both a rat acute carrageenan-induced arthritis model and rat chronic adjuvant induced arthritis model for arthritic inflammation. UCX® anti-inflammatory activity was then monitored over time.ResultsUCX® cells stained positive for CD44, CD73, CD90 and CD105; and negative for CD14, CD19 CD31, CD34, CD45 and HLA-DR; and were capable to differentiate into adipocyte, chondrocyte and osteoblast-like cells. UCX® cells were shown to repress T-cell activation and promote the expansion of Tregs better than bone marrow mesenchymal stem cells (BM-MSCs). Accordingly, xenogeneic UCX® administration in an acute carrageenan-induced arthritis model showed that human UCX® cells can reduce paw edema in vivo more efficiently than BM-MSCs. Finally, in a chronic adjuvant induced arthritis model, animals treated with intra-articular (i.a.) and intra-peritoneal (i.p.) infusions of UCX® cells showed faster remission of local and systemic arthritic manifestations.ConclusionThe results suggest that UCX® cells may be an effective and promising new approach for treating both local and systemic manifestations of inflammatory arthritis.