scholarly journals Lower retention after retrograde coronary venous infusion compared with intracoronary infusion of mesenchymal stromal cells in the infarcted porcine myocardium

2019 ◽  
Vol 3 (1) ◽  
pp. e000006 ◽  
Author(s):  
Wouter A Gathier ◽  
Mira van der Naald ◽  
Bas R van Klarenbosch ◽  
Anton E Tuinenburg ◽  
John LM Bemelmans ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Ex Vivo ◽  
Pericardial Fluid ◽  
Cardiac Cell ◽  
Cell Retention ◽  
Intramyocardial Injection ◽  
Advantages And Disadvantages ◽  
Venous Infusion

BackgroundCommonly used strategies for cell delivery to the heart are intramyocardial injection and intracoronary (IC) infusion, both having their advantages and disadvantages. Therefore, alternative strategies, such as retrograde coronary venous infusion (RCVI), are explored. The aim of this confirmatory study was to compare cardiac cell retention between RCVI and IC infusion. As a secondary end point, the procedural safety of RCVI is assessed.MethodsFour weeks after myocardial infarction, 12 pigs were randomised to receive mesenchymal stromal cells, labelled with Indium-111, via RCVI (n=6) or IC infusion (n=6). Four hours after cell administration, nuclear imaging was performed to determine the number of cells retained in the heart both in vivo and ex vivo. Procedure-related safety measures were reported.ResultsCardiac cell retention is significantly lower after RCVI compared with IC infusion (in vivo: RCVI: median 2.89% vs IC: median 13.74%, p=0.002, ex vivo: RCVI: median 2.55% vs IC: median 39.40%, p=0.002). RCVI led to development of pericardial fluid and haematomas on the frontal wall of the heart in three cases. Coronary venous dissection after RCVI was seen in three pigs, of which one also developed pericardial fluid and a haematoma. IC infusion led to no flow in one pig.ConclusionRCVI is significantly less efficient in delivering cells to the heart compared with IC infusion. RCVI led to more procedure-related safety issues than IC infusion, with multiple cases of venous dissection and development of haematomas and pericardial fluid collections.

Hypoxic Preconditioning of Mesenchymal Stromal Cells Induces Metabolic Changes, Enhances Survival, and Promotes Cell Retention In Vivo

Stem Cells ◽  
2015 ◽  
Vol 33 (6) ◽  
pp. 1818-1828 ◽  
Author(s):  
Julie Beegle ◽  
Kinga Lakatos ◽  
Stefanos Kalomoiris ◽  
Heather Stewart ◽  
R. Rivkah Isseroff ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Metabolic Changes ◽  
Hypoxic Preconditioning ◽  
Cell Retention

scholarly journals Mesenchymal stromal cells release CXCL1/2/8 and induce chemoresistance and macrophage polarization

2018 ◽  
Author(s):  
Bettina Couderc ◽  
Augustin Le Naour ◽  
Mélissa Prat ◽  
Renaud Mevel ◽  
Benoit Thibault ◽  
...  
Keyword(s):  
Tumor Progression ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Therapeutic Strategy ◽  
Ex Vivo ◽  
Macrophage Polarization ◽  
Activated Macrophages ◽  
High Concentrations ◽  
Ovarian Tumor Cells

Factors released by surrounding cells such as cancer-associated mesenchymal stromal cells (CA-MSCs) are involved in tumor progression and chemoresistance. We determine the mechanisms by which a naïve MSC could become a CA-MSC and characterize CA-MSCs. Ovarian tumor cells (OTC) trigger the transformation of MSCs to CA-MSCs expressing different pro-tumoral, genes and secreting high amounts of CXCR1/2 ligands (CXCL1, CXCL2 and IL-8) implicated in the chemoresistance of cancer cells. CXCR1/2 ligands can also inhibit the immune response against OTC. Indeed, through their released factors, CA-MSCs can trigger the differentiation of monocytes to pro-tumoral M2 phenotype macrophages known to promote the tumor progression. When CXCR1/2 receptors are inhibited, these CA-MSC-activated macrophages lose their M2 functions and acquire an anti-tumoral phenotype. Both ex vivo and in vivo a CXCR1/2 inhibitor can sensitize OTC to carboplatin even in the presence of a pro-tumoral microenvironment. This inhibitor can circumvent the pro-tumoral effects of CA-MSCs. As high concentrations of CXCR1/2 ligands in blood from patients can be associated with chemoresistance, CXCR1/2 inhibition could be a potential therapeutic strategy to revert chemoresistance.

scholarly journals Ex vivo Ikkβ ablation rescues the immunopotency of mesenchymal stromal cells from diabetics with advanced atherosclerosis

2020 ◽  
Author(s):  
Ozge Kizilay Mancini ◽  
David N Huynh ◽  
Liliane Menard ◽  
Dominique Shum-Tim ◽  
Huy Ong ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Inflammatory Responses ◽  
Ex Vivo ◽  
Therapeutic Effects ◽  
Atherosclerotic Cardiovascular Disease ◽  
Myocardial Repair ◽  
Iκb Kinase

Abstract Aims Diabetes is a conventional risk factor for atherosclerotic cardiovascular disease and myocardial infarction (MI) is the most common cause of death among these patients. Mesenchymal stromal cells (MSCs) in patients with type 2 diabetes mellitus (T2DM) and atherosclerosis have impaired ability to suppress activated T-cells (i.e. reduced immunopotency). This is mediated by an inflammatory shift in MSC-secreted soluble factors (i.e. pro-inflammatory secretome) and can contribute to the reduced therapeutic effects of autologous T2DM and atherosclerosis-MSC post-MI. The signalling pathways driving the altered secretome of atherosclerosis- and T2DM-MSC are unknown. Specifically, the effect of IκB kinase β (IKKβ) modulation, a key regulator of inflammatory responses, on the immunopotency of MSCs from T2DM patients with advanced atherosclerosis has not been studied. Methods and results MSCs were isolated from adipose tissue obtained from patients with (i) atherosclerosis and T2DM (atherosclerosis+T2DM MSCs, n = 17) and (ii) atherosclerosis without T2DM (atherosclerosis MSCs, n = 17). MSCs from atherosclerosis+T2DM individuals displayed an inflammatory senescent phenotype and constitutively expressed active forms of effectors of the canonical IKKβ nuclear factor-κB transcription factors inflammatory pathway. Importantly, this constitutive pro-inflammatory IKKβ signature resulted in an altered secretome and impaired in vitro immunopotency and in vivo healing capacity in an acute MI model. Notably, treatment with a selective IKKβ inhibitor or IKKβ knockdown (KD) (clustered regularly interspaced short palindromic repeats/Cas9-mediated IKKβ KD) in atherosclerosis+T2DM MSCs reduced the production of pro-inflammatory secretome, increased survival, and rescued their immunopotency both in vitro and in vivo. Conclusions Constitutively active IKKβ reduces the immunopotency of atherosclerosis+T2DM MSC by changing their secretome composition. Modulation of IKKβ in atherosclerosis+T2DM MSCs enhances their myocardial repair ability.

Ex Vivo Gene Therapy for Haemophilia A: FVIII-Overexpressing Mesenchymal Stromal Cells Fail To Produce Sustained Therapeutic Levels of Circulating FVIII In Vivo.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 5472-5472
Author(s):  
Daniel L. Coutu ◽  
Moira Francois ◽  
Jessica Cuerquis ◽  
Hideto Matsui ◽  
David Lillicrap ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Lentiviral Vector ◽  
Ex Vivo ◽  
Coagulation Factor ◽  
Scid Mice ◽  
Limiting Factor ◽  
Haemophilia A

Abstract The use of ex vivo transfected autologous fibroblasts was explored for the delivery of coagulation factor VIII (FVIII) to treat haemophilia A patients, albeit with mitigated results (Roth et al., NEJM 2001). A limiting factor to this approach is the limited expansion potential of somatic cells and cellular dose manufactured for therapeutic use. The use of adult stem or progenitor cells offers a possible remedy to this specific challenge because of their high proliferative capacities, allowing the generation of large number of cells ex vivo. Mesenchymal stromal cells (MSCs) can easily be isolated by bone marrow aspiration and extensively expanded ex vivo using standard tissue culture procedures. They also possess endogenous wound healing properties and are relatively easy to gene engineer. When engineered to secrete therapeutic plasma-soluble proteins, they could be injected with a supporting matrix - as a subcutaneous plug - and used to treat a variety of haematological disorders responsive to secreted proteins such as haemophilia A and B and anaemia for instance. The retrievability of the implanted cells is desirable if unexpected side effects were to occur. The aim of this project was to test a gene engineered MSC platform to treat a clinically relevant model disease, haemophilia A. Canine MSCs (cMSCs) isolated from normal and haemophilia A dogs were genetically modified using a 3rd generation lentiviral vector carrying the B-domain deleted canine FVIII (cFVIII) gene. In vitro production of cFVIII was assessed by ELISA and found to be about 2,4 U/106 cells/24h. 107 of those cells were injected intraperitoneally in NOD-SCID mice and transient expression (8–10 days) of therapeutic levels of cFVIII (up to 10%) could be detected using an ELISA capable of discriminating cFVIII from murine FVIII. However, when these cells were embedded in an injectable human compatible collagen based scaffolding material (Contigen) and implanted subcutaneously into NOD-SCID mice no cFVIII could be detected above background in any animals. Because we previously demonstrated that erythropoietin gene-modified MSCs implanted as described led to an increase in hematocrit for an extended period (Eliopoulos et al. Mol Ther 2004), we believe that the failure of the present approach is attributable to idiosyncrasies of the FVIII system. Of particular relevance is the requirement for the presence of von Willebrand factor (vWF) in stabilizing the FVIII protein and protect it from degradation. We already have evidence that ex vivo transduced endothelial progenitor cells known to produce endogenous vWF can serve as better delivery vehicles for FVIII as part of a matrix embedded subcutaneous plug and lead to prolonged circulating levels of FVIII in mice (Matsui et al., submitted). These data support the hypothesis that FVIII delivery by engineered somatic and stem cells likely requires co-expression of key cofactors, such as vWF, for durable and clinically relevant effects in vivo.

scholarly journals Tumor cells educate mesenchymal stromal cells to release chemoprotective and immunomodulatory factors

2019 ◽  
Vol 12 (3) ◽  
pp. 202-215 ◽  
Author(s):  
Augustin Le Naour ◽  
Mélissa Prat ◽  
Benoît Thibault ◽  
Renaud Mével ◽  
Léa Lemaitre ◽  
...  
Keyword(s):  
Tumor Progression ◽  
Tumor Cells ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Chemokine Receptors ◽  
Ovarian Tumor ◽  
Ex Vivo ◽  
High Concentrations ◽  
Ovarian Tumor Cells

Abstract Factors released by surrounding cells such as cancer-associated mesenchymal stromal cells (CA-MSCs) are involved in tumor progression and chemoresistance. In this study, we characterize the mechanisms by which naïve mesenchymal stromal cells (MSCs) can acquire a CA-MSCs phenotype. Ovarian tumor cells trigger the transformation of MSCs to CA-MSCs by expressing pro-tumoral genes implicated in the chemoresistance of cancer cells, resulting in the secretion of high levels of CXC chemokine receptors 1 and 2 (CXCR1/2) ligands such as chemokine (C-X-C motif) ligand 1 (CXCL1), CXCL2, and interleukin 8 (IL-8). CXCR1/2 ligands can also inhibit the immune response against ovarian tumor cells. Indeed, through their released factors, CA-MSCs promote the differentiation of monocytes towards M2 macrophages, which favors tumor progression. When CXCR1/2 receptors are inhibited, these CA-MSC-activated macrophages lose their M2 properties and acquire an anti-tumoral phenotype. Both ex vivo and in vivo, we used a CXCR1/2 inhibitor to sensitize ovarian tumor cells to carboplatin and circumvent the pro-tumoral effects of CA-MSCs. Since high concentrations of CXCR1/2 ligands in patients’ blood are associated with chemoresistance, CXCR1/2 inhibition could be a potential therapeutic strategy to revert carboplatin resistance.

scholarly journals Differential effects of the cystic fibrosis lung inflammatory environment on mesenchymal stromal cells

2020 ◽  
Vol 319 (6) ◽  
pp. L908-L925
Author(s):  
Soraia C. Abreu ◽  
Thomas H. Hampton ◽  
Evan Hoffman ◽  
Jacob Dearborn ◽  
Alix Ashare ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cystic Fibrosis ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Ex Vivo ◽  
Lavage Fluid ◽  
Cell Therapies ◽  
Acute Exacerbations ◽  
Aspergillus Sp

Growing evidence demonstrates that human mesenchymal stromal cells (MSCs) modify their in vivo anti-inflammatory actions depending on the specific inflammatory environment encountered. Understanding this better is crucial to refine MSC-based cell therapies for lung and other diseases. Using acute exacerbations of cystic fibrosis (CF) lung disease as a model, the effects of ex vivo MSC exposure to clinical bronchoalveolar lavage fluid (BALF) samples, as a surrogate for the in vivo clinical lung environment, on MSC viability, gene expression, secreted cytokines, and mitochondrial function were compared with effects of BALF collected from healthy volunteers. CF BALF samples that cultured positive for Aspergillus sp. (Asp) induced rapid MSC death, usually within several hours of exposure. Further analyses suggested the fungal toxin gliotoxin as a potential mediator contributing to CF BALF-induced MSC death. RNA sequencing analyses of MSCs exposed to either Asp+ or Asp− CF BALF samples identified a number of differentially expressed transcripts, including those involved in interferon signaling, antimicrobial gene expression, and cell death. Toxicity did not correlate with bacterial lung infections. These results suggest that the potential use of MSC-based cell therapies for CF or other lung diseases may not be warranted in the presence of Aspergillus.

scholarly journals Mobilisation of Joint-resident Mesenchymal Stromal Cells for Articular Cartilage Regeneration

2017 ◽  
Vol 5 (2_suppl2) ◽  
pp. 2325967117S0007
Author(s):  
Alam Khalil Khan ◽  
Thomas Baboolal ◽  
Owen Wall ◽  
Elena Jone ◽  
Dennis Mcgonagle
Keyword(s):  
Synovial Fluid ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Ex Vivo ◽  
Platelet Rich Plasma ◽  
Blood Clot ◽  
Cartilage Regeneration ◽  
Mechanical Agitation ◽  
Cytology Brush

Background and Objectives: Microfracture is a recognized procedure used to treat isolated cartilage injuries or defects, in which bone marrow mesenchymal stromal cells (BM-MSCs) are thought to migrate into the resulting blood clot, leading to subsequent cartilage repair via fibrocartilage formation. The discovery of MSCs in the synovium and synovial fluid (SF) provides a potential mechanism for repairing cartilage from the top down via their migration and homing to the microfracture site, however SF-MSCs low in number and usually lost with joint irrigation. The purpose of this work was threefold; first to test the hypothesis that SF-MSCs can be replaced, and also their numbers further increased by synovial agitation, second that these cells were capable of rapid adhesion to clots and third that the clot composition improve MSC migration. Materials-Methods: Ex-vivo mechanical agitation of human superficial synovium and in vivo intra-operative agitation of synovium in patients undergoing arthroscopy were performed using a Cytology brush and custom designed Synovial brush. Colony-forming unit-fibroblast (CFU-F) assay was performed to quantify released MSCs. Adhesion to clots was studied by comparing Platelet Rich Plasma (PRP), Whole Blood (WB) and Fibrin Glue (FG). Migration studies were performed using passage 2-4 synovial MSCs in trans-well migration assay. MSC migration, over a five hour period, was compared between PRP and pooled human Platelet Lysate (hPL). Results: Ex-vivo mechanical agitating of the synovium with the cytology brush compared to irrigation alone increased MSC number 2.7-fold (n=10, p=0.002). Irrigation during arthroscopy was seen to effectively remove the majority of CFU-Fs for the synovial fluid. Use of a custom designed synovial brush, compared to the cytology brush resulted in a median 65-fold increase in the number of CFU-Fs (n=8, p=0.0148). Trilineage differentiation of released synovial MSCs was at least comparable to donor match synovial fluid MSCs. These MSCs adhered to clots within 30 minutes with no difference seen between clot compositions. Released synovial MSCs demonstrated a trend for a better migration towards hPL compared to PRP. Conclusions: Existing surgical procedures wash away SF-MSCs. Using a novel brushing technique and a custom designed synovial brush, synovial MSCs can be mechanically released in vivo, and these cells were capable of migration and rapid adhesion to clots. Collectively these findings aid in the rapid replenishment of endogenous minimally manipulated MSCs, and provide a one-stage procedure combining synovial brushing with microfracture, as a strategy for cost effective joint repair.

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