Evaluation of Neuroprotective And Immunomodulatory Properties of Mesenchymal Stem Cells in An Ex-vivo Retinal Explant Model

Abstract Background: Mesenchymal stem cells (MSCs) have raised considerable hope for the treatment of glaucoma. Their neuroprotective and regenerative potentials are particularly interesting for this degenerative neuropathy in which retinal ganglion cell (RGC) death leads to a progressive loss of visual field and eventually vision. Yet, despite promising results in animal models, no definitive treatment has been developed, and safety concerns have been reported in human trials. Microglial immunomodulation represents a promising therapeutic approach in which MSCs might play a crucial role. In the present study, we investigated the neuroprotective and immunomodulatory properties as well as the safety of MSCs in an ex vivo neuroretina explant model.Methods: Labeled rat bone marrow MSCs were placed in co-culture with rat retinal explants after optic nerve axotomy. We analyzed the neuroprotective effect of MSCs on RGC survival by immunofluorescence using RBPMS, Brn3a and NeuN markers. Gliosis and retinal microglial activation were measured using GFAP, CD68 and ITGAM mRNA quantification and GFAP, CD68 and Iba1 immunofluorescence staining. We also analyzed the mRNA expression of both ‘M1’ or classically activated state inflammatory cytokines (TNFα, IL1β and IL6), and ‘M2’ or alternatively activated state microglial markers (Arginase 1, IL10, CD163, and TNFAIP6).Results: The number of RGCs was significantly higher in retinal explants cocultured with MSCs compared to the control group at Day 7 following optic nerve axotomy. Retinal explants co-cultured with MSCs showed decreased mRNA markers of gliosis and microglial activation, and immunostaining revealed that GFAP, Iba1 and CD68 were limited to the internal layers of the retina compared to controls showing expression of activated microglia throughout the retina. In addition, MSCs inhibited the M1 phenotype of the microglia. However, edema of the explants was observed in the MSC co-culture group, with an increase of fibronectin labelling at the surface of the explant corresponding to an epiretinal membrane-like phenotype. Conclusion: Using an ex vivo model, we demonstrated a neuroprotective and immunomodulatory effect of MSCs on RGCs. Unfortunately, the presence of MSCs also led to explant edema and epiretinal membrane formation, as described in human trials. Using the MSC secretome might offer the beneficial effects of MSCs without their potential adverse effects, through paracrine signaling.

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Immunogenicity and immunomodulatory properties of umbilical cord lining mesenchymal stem cells for regenerative therapies

The Thoracic and Cardiovascular Surgeon ◽

10.1055/s-0030-1268919 ◽

2011 ◽

Vol 59 (S 01) ◽

Author(s):

M Stubbendorff ◽

T Deuse ◽

K Tang-Quan ◽

H Reichenspurner ◽

RC Robbins ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Umbilical Cord ◽

Immunomodulatory Properties ◽

Regenerative Therapies

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Comparative analysis of the immunomodulatory properties of equine adult-derived mesenchymal stem cells

Cell Medicine ◽

10.3727/215517911x647217 ◽

2012 ◽

Author(s):

Danielle D. Carrade ◽

Michael W. Lame ◽

Michael S. Kent ◽

Kaitlin C. Clark ◽

Naomi J. Walker ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Comparative Analysis ◽

Immunomodulatory Properties

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130 Burn Injury Reduces Bone Marrow Mesenchymal Stem Cells and Sensitizes Their Adrenergic Receptor Subgroups in a Murine Model

Journal of Burn Care & Research ◽

10.1093/jbcr/irab032.134 ◽

2021 ◽

Vol 42 (Supplement_1) ◽

pp. S87-S88

Author(s):

Kuzhali Muthumalaiappan ◽

Maria Camargo Johnson ◽

Julia Walczak ◽

Vimal Subramaniam ◽

Anthony J Baldea ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Adrenergic Receptor ◽

Burn Injury ◽

Bone Marrow Cells ◽

Ex Vivo ◽

Traumatic Injury ◽

Hematopoietic Cell ◽

The Right

Abstract Introduction Previous burn and traumatic injury studies have established that adrenergic signaling is increased after burn injury and may lead to an impairment of hematopoietic cell development in the bone marrow (BM). Nonetheless, mesenchymal stem cells (MSCs), which have gained momentum in regenerative medicine also play a predominant role in the BM niche. Understanding the propensity of the adrenergic receptor (AR) response by MSCs can be utilized for devising targeted therapies. However, the traditional plastic adherence procedure using ex vivo culture of BM cells for several weeks may skew the actual characteristics of MSCs. Our current study focused on isolating MSCs from freshly obtained BM in a murine scald burn model with a goal to characterize the expression pattern of native AR subgroups present on BM MSCs as compared to sham mice. Methods Eight, two-month-old adult female mice were subjected to a 15% total body 3rd degree burn or sham burn. The mice were sacrificed 7 days later. Femurs were removed and total bone marrow cells were flushed out. Multi parametric flow cytometry was used to gate for cells negative for hematopoietic cell markers (CD45, CD11B) and positive for MSC markers (CD105, CD106, SSEA, Ly6A) and AR subgroups (α1, α2, β1, β2, β3). We measured the number of BM MSCs, quantified the subtypes of ARs present on MSCs, and compared the ratio of AR antibody binding per total MSC population. Results Overall the frequency of MSCs per million total BM cells decreased by 48% post-burn injury with165,300 ± 194 in sham versus 110,000 ± 30 in burn displayed as bar graph in Panel A. Over 90% of MSCs consistently express β2 AR and only 10% express α2 AR subgroup in both scald and sham burn. Presence of other subgroups ranged from 50% to 80% of MSCs as seen in histograms to the right of dotted line in Panel B. Our AR propensity score based on AR mean fluorescence intensity adjusted to total number of MSCs present was increased by 2.8-fold for α1, 2.5-fold for β1, 1.6-fold for β3, and 1.3-fold for β2 AR subgroups (Panel C). These findings indicate burn injury not only decreases the frequency of BM MSCs but also increases the affinity of certain AR subgroups present on MSCs. Since BM MSCs are the major source of cytokines, chemokines and growth factors; detailed studies on AR mediated signaling in BM MSCs is warranted. Conclusions Polarization of AR signaling in BM MSCs by burn-induced catecholamines may have broader implications for comorbidities such as bone resorption and muscle wasting observed in human patients post burn trauma.

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Effects of continuous passage on the immunomodulatory properties of equine bone marrow-derived mesenchymal stem cells in vitro

Veterinary Immunology and Immunopathology ◽

10.1016/j.vetimm.2021.110203 ◽

2021 ◽

Vol 234 ◽

pp. 110203

Author(s):

Shannon S. Connard ◽

Renata L. Linardi ◽

Kayla M. Even ◽

Alix K. Berglund ◽

Lauren V. Schnabel ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Immunomodulatory Properties ◽

Equine Bone

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L-Glutamine in vitro Modulates some Immunomodulatory Properties of Bone Marrow Mesenchymal Stem Cells

Stem Cell Reviews and Reports ◽

10.1007/s12015-017-9746-0 ◽

2017 ◽

Vol 13 (4) ◽

pp. 482-490 ◽

Cited By ~ 10

Author(s):

Guilherme Galvão dos Santos ◽

Araceli Aparecida Hastreiter ◽

Talita Sartori ◽

Primavera Borelli ◽

Ricardo Ambrósio Fock

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Marrow Mesenchymal Stem Cells ◽

Immunomodulatory Properties

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DNA Methylation Changes duringIn VitroPropagation of Human Mesenchymal Stem Cells: Implications for Their Genomic Stability?

Stem Cells International ◽

10.1155/2013/192425 ◽

2013 ◽

Vol 2013 ◽

pp. 1-9 ◽

Cited By ~ 33

Author(s):

Angela Bentivegna ◽

Mariarosaria Miloso ◽

Gabriele Riva ◽

Dana Foudah ◽

Valentina Butta ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Ex Vivo ◽

Genomic Stability ◽

Great Promise ◽

Human Mscs ◽

Low Oxygen ◽

Long Term Culture ◽

Functional Changes

Mesenchymal stem cells (MSCs) hold great promise for the treatment of numerous diseases. A major problem for MSC therapeutic use is represented by the very low amount of MSCs which can be isolated from different tissues; thusex vivoexpansion is indispensable. Long-term culture, however, is associated with extensive morphological and functional changes of MSCs. In addition, the concern that they may accumulate stochastic mutations which lead the risk of malignant transformation still remains. Overall, the genome of human MSCs (hMSCs) appears to be apparently stable throughout culture, though transient clonal aneuploidies have been detected. Particular attention should be given to the use of low-oxygen environment in order to increase the proliferative capacity of hMSCs, since data on the effect of hypoxic culture conditions on genomic stability are few and contradictory. Furthermore, specific and reproducible epigenetic changes were acquired by hMSCs duringex vivoexpansion, which may be connected and trigger all the biological changes observed. In this review we address current issues on long-term culture of hMSCs with a 360-degree view, starting from the genomic profiles and back, looking for an epigenetic interpretation of their genetic stability.

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Mesenchymal stem cells accelerated growth and metastasis of neuroblastoma and preferentially homed towards both primary and metastatic loci in orthotopic neuroblastoma model

BMC Cancer ◽

10.1186/s12885-021-08090-2 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Jiao-Le Yu ◽

Shing Chan ◽

Marcus Kwong-Lam Fung ◽

Godfrey Chi-Fung Chan

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Primary Tumor ◽

Imaging System ◽

Ex Vivo ◽

Tumor Formation ◽

Human Neuroblastoma Cell ◽

Fluorescence Signal ◽

Human Neuroblastoma

Abstract Background Majority of neuroblastoma patients develop metastatic disease at diagnosis and their prognosis is poor with current therapeutic approach. Major challenges are how to tackle the mechanisms responsible for tumorigenesis and metastasis. Human mesenchymal stem cells (hMSCs) may be actively involved in the constitution of cancer microenvironment. Methods An orthotopic neuroblastoma murine model was utilized to mimic the clinical scenario. Human neuroblastoma cell line SK-N-LP was transfected with luciferase gene, which were inoculated with/without hMSCs into the adrenal area of SCID-beige mice. The growth and metastasis of neuroblastoma was observed by using Xenogen IVIS 100 in vivo imaging and evaluating gross tumors ex vivo. The homing of hMSCs towards tumor was analyzed by tracing fluorescence signal tagged on hMSCs using CRI Maestro™ imaging system. Results hMSCs mixed with neuroblastoma cells significantly accelerated tumor growth and apparently enhanced metastasis of neuroblastoma in vivo. hMSCs could be recruited by primary tumor and also become part of the tumor microenvironment in the metastatic lesion. The metastatic potential was consistently reduced in lung and tumor when hMSCs were pre-treated with stromal cell derived factor-1 (SDF-1) blocker, AMD3100, suggesting that the SDF-1/CXCR4 axis was one of the prime movers in the metastatic process. Conclusions hMSCs accelerated and facilitated tumor formation, growth and metastasis. Furthermore, the homing propensity of hMSCs towards both primary tumor and metastatic loci can also provide new therapeutic insights in utilizing bio-engineered hMSCs as vehicles for targeted anti-cancer therapy.

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