Characterization and Comparison of Human and Ovine Mesenchymal Stromal Cells from Three Corresponding Sources

Currently, there is an increasing focus on mesenchymal stromal cells (MSC) as therapeutic option in bone pathologies as well as in general regenerative medicine. Although human MSCs have been extensively characterized and standardized, ovine MSCs are poorly understood. This limitation hampers clinical progress, as sheep are an excellent large animal model for orthopedic studies. Our report describes a direct comparison of human and ovine MSCs from three corresponding sources under the same conditions. All MSCs presented solid growth behavior and potent immunomodulatory capacities. Additionally, we were able to identify common positive (CD29, CD44, CD73, CD90, CD105, CD166) and negative (CD14, CD34, CD45, HLA-DR) surface markers. Although both human and ovine MSCs showed strong osteogenic potential, direct comparison revealed a slower mineralization process in ovine MSCs. Regarding gene expression level, both human and ovine MSCs presented a comparable up-regulation of Runx2 and a trend toward down-regulation of Col1A during osteogenic differentiation. In summary, this side by side comparison defined phenotypic similarities and differences of human and ovine MSCs from three different sources, thereby contributing to a better characterization and standardization of ovine MSCs. The key findings shown in this report demonstrate the utility of ovine MSCs in preclinical studies for MSC-based therapies.

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Quantitative Efficacy and Fate of Mesenchymal Stromal Cells Targeted to Cardiac Sites by Radiofrequency Catheter Ablation

Cell Transplantation ◽

10.1177/0963689720914236 ◽

2020 ◽

Vol 29 ◽

pp. 096368972091423

Author(s):

Rizwan Malik ◽

Fabrice F. Darche ◽

Rasmus Rivinius ◽

Anja Seckinger ◽

Ulf Krause ◽

...

Keyword(s):

Stem Cell ◽

Catheter Ablation ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Radiofrequency Catheter Ablation ◽

Functional Integration ◽

Right Atrial ◽

Blue Staining ◽

Large Animal ◽

Human Mscs

Engraftment and functional integration of stem cells or stem cell-derived cells within cardiac tissue is an important prerequisite for cell replacement therapy aiming at the treatment of heart disease. Recently, a novel intravenous approach for application of mesenchymal stromal cells (MSCs) to cardiac sites has been established using radiofrequency catheter ablation (RFCA)-guided targeting, bypassing the need for open chest surgery or direct myocardial cell injection. However, little is known about the quantitative efficacy and longevity of this strategy. We performed selective power-controlled RFCA with eight ablation pulses (30 W, 60 s each) to induce heat-mediated lesions at the right atrial appendices (RAAs) of pigs. Different concentrations of human bone marrow-derived MSCs (105 to 1.6 × 106 cells/kg bodyweight) labeled with superparamagnetic iron oxide (SPIO) particles were infused intravenously in nine pigs one d after RFCA treatment and hearts were explanted 8 d later to quantify the number of engrafted cells. Prussian blue staining revealed high numbers of SPIO-labeled cells in areas surrounding the RFCA-induced lesions. Cell numbers were evaluated by quantitative real-time polymerase chain reaction using specific primers for human MSCs (hMSCs), which indicated that up to 106 hMSCs, corresponding to ∼3.9% of the systemically applied human cells, engrafted within the RAAs of RFCA-treated pigs. Of note, infused hMSCs were observed in nontargeted organs, as well, but appeared at very low concentrations. To assess long-term deposition of MSCs, RAAs of three pigs were analyzed after 6 months, which revealed few persisting hMSCs at targeted sites. RFCA-mediated targeting of MSCs provides a novel minimal invasive strategy for cardiac stem cell engraftment. Qualitative and quantitative results of our large animal experiments indicate an efficient guidance of MSCs to selected cardiac regions, although only few cells remained at targeted sites 6 mo after cell transplantation.

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CD14 dictates differential activation of mesenchymal stromal cells through AKT, NF-κB and P38 signals

Bioscience Reports ◽

10.1042/bsr20190807 ◽

2019 ◽

Vol 39 (7) ◽

Cited By ~ 5

Author(s):

Menghui Jiang ◽

Tianlin Gao ◽

Yuansheng Liu ◽

Xue Cao ◽

Yanting Li ◽

...

Keyword(s):

Signaling Pathway ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Expression Patterns ◽

Mrna Level ◽

Human Mscs ◽

Differentiation Potential ◽

Tlr4 Signaling ◽

Tlr4 Signaling Pathway ◽

Different Sources

Abstract Mesenchymal stromal cells (MSCs) widely exist in many tissues and have multiple differentiation potential and immunomodulatory capacities. Recently, MSCs have become promising tools for the treatment of various degenerative disorders and autoimmune diseases. The properties of MSCs could be modified in different microenvironments. Thus, it is important to explore the factors controlling MSC function. The presence of Toll-like receptors (TLRs) in MSCs was demonstrated according to previous studies. Consistently, we also illustrated the expression of TLRs in both murine and human MSCs, and displayed that the expression patterns of TLRs in MSCs from different sources. Furthermore, we explored the role of TLR and TLR signaling pathway in MSCs. Interestingly, activation of TLR4-induced expression of cytokines and some specific genes in MSCs. However, MSCs retained much lower mRNA level compared with macrophages. We explored the expression of CD14 in MSCs from different sources, which played a vital role in TLR4 signaling pathway, and found that MSCs are almost negative for CD14. Moreover, only partial activation of TLR4 signaling pathway was observed in MSCs, with no activation of AKT, NF-κB and P38. Here, in the study we defined TLR expression, function and activation in MSCs, which is critical for designing MSC-based therapies.

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Persistence and proliferation of human mesenchymal stromal cells in the right ventricular myocardium after intracoronary injection in a large animal model of pulmonary hypertension

Cytotherapy ◽

10.1016/j.jcyt.2017.03.002 ◽

2017 ◽

Vol 19 (6) ◽

pp. 668-679 ◽

Cited By ~ 5

Author(s):

Roza Badr Eslam ◽

Kevin Croce ◽

Fernanda Marinho Mangione ◽

Robert Musmann ◽

Jane A. Leopold ◽

...

Keyword(s):

Animal Model ◽

Pulmonary Hypertension ◽

Right Ventricular ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Ventricular Myocardium ◽

Large Animal Model ◽

Large Animal ◽

Human Mesenchymal Stromal Cells ◽

The Right

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In Vivo Osteogenic Potential of Mesenchymal Stromal Cells Derived from Patient''s Bone Marrow on Hydroxyapatite Ceramics

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.330-332.1157 ◽

2007 ◽

Vol 330-332 ◽

pp. 1157-1160 ◽

Cited By ~ 1

Author(s):

Asako Matsushima ◽

Noriko Kotobuki ◽

Hiroko Machida ◽

Toru Morishita ◽

Yoshinori Takakura ◽

...

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Bone Matrix ◽

Osteogenic Potential ◽

Human Mscs ◽

Hard Tissue ◽

Culture Dish ◽

Hydroxyapatite Ceramics

Since 2001, we have started tissue engineered approach for hard tissue repair using mesenchymal stromal cells (MSCs) derived from patient’s bone marrow. MSCs were culture expanded on culture dish, then applied on various ceramics including hydroxyapatite (HA) ceramics. The MSCs on the ceramics were further cultured in osteogenic media to induce osteognenic differentiation. The differentiation resulted in appearance of bone forming osteoblasts as well as bone matrix on the ceramics, thus we could fabricate the tissue engineered bone. We have reported that the tissue engineered bone is effective for treatment of large bone defect, which is difficult to repair only with artificial materials such as HA ceramics. The present study focused on osteogenic capability of cryopreserved human MSCs derived from patients who already were treated by the tissue engineered bone. The MSCs showed high alkaline phosphatase activity together with abundant bone matrix formation when cultured in osteogenic media. The MSCs also showed in vivo new bone formation when implanted at subcutaneous sites of athymic nude rats. Based on the results, we concluded that the tissue engineering approach is a reliable method to be used in hard tissue regeneration.

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Long-Term Cell Tracking following Local Injection of Mesenchymal Stromal Cells in the Equine Model of Induced Tendon Disease

Cell Transplantation ◽

10.3727/096368916x692104 ◽

2016 ◽

Vol 25 (12) ◽

pp. 2199-2211 ◽

Cited By ~ 24

Author(s):

Janina Burk ◽

Dagmar Berner ◽

Walter Brehm ◽

Aline Hillmann ◽

Carolin Horstmeier ◽

...

Keyword(s):

Animal Model ◽

Mesenchymal Stromal Cells ◽

Rhodamine B ◽

Stromal Cells ◽

Cell Tracking ◽

Local Application ◽

Large Animal Model ◽

Large Animal

Tendon disease has been treated with multipotent mesenchymal stromal cells (MSCs) in the equine large-animal model with promising success. The aim of this study was to gain more insight into the fate and biodistribution of MSCs after local application into tendon lesions by long-term cell tracking in this large-animal model. Superficial digital flexor tendon lesions were induced in all limbs in six horses and injected with 10 × 10 6 Molday ION Rhodamine B™-labeled MSCs suspended in serum or serum alone. Follow-up was performed using low-field magnetic resonance imaging (MRI), flow cytometry, and histology. Cell tracking based on the hypointense artifacts induced by the superparamagnetic iron oxide (SPIO) labeling agent in MRI as well as based on Rhodamine B fluorescence was feasible. However, Prussian blue staining for assessment of histology was not entirely specific for SPIO. Labeled cells could be traced at their injection site by MRI as well as histology for the whole follow-up period of 24 weeks. Although the numbers of labeled cells within the injected tendon lesions decreased over time, part of the applied cells appeared to remain viable and integrated within the injured tissue. Furthermore, small numbers of labeled cells were identified in peripheral blood within the first 24 h after cell injection and could also be found until week 24 within the contralateral control tendon lesions that had been injected with serum. The present findings unveil details on MSC biodistribution and persistence after their local application, which are of clinical relevance with regard to MSC safety and mechanisms of action.

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Evaluation of Potential Application of Wharton’s Jelly-Derived Human Mesenchymal Stromal Cells and its Conditioned Media for Dermal Regeneration using Rat Wound Healing Model

Cells Tissues Organs ◽

10.1159/000513895 ◽

2021 ◽

pp. 1-14

Author(s):

Caroline Mathen ◽

Mrunal Ghag Sawant ◽

Raghubansh Gupta ◽

Wilfrid Dsouza ◽

Shilpa G. Krishna

Keyword(s):

Wound Healing ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Therapeutic Potential ◽

Wound Care ◽

Free Cell ◽

Conditioned Media ◽

Human Umbilical Cord ◽

Human Mscs ◽

Wound Model

Mesenchymal stromal cells and the derived conditioned media represent an area of tremendous medical interest and, among other clinical applications, are currently being extensively explored for wound healing. The aim of this study was to comparatively evaluate the wound healing potential of xeno-free human umbilical cord-derived mesenchymal stromal cells (MSCs) and the conditioned media (CM) in a full-thickness excision wound model in rats. The evaluation parameters included rate of wound healing, serum cytokine analyses, collagen content, histopathology, and hyperspectral imaging as an independent qualitative and quantitative tool. Both the cell-based and cell-free approaches scored better in lower inflammation, as evidenced in lower IL-10 and stable IL-6 levels, and improved rate of wound healing (<i>p</i> < 0.0001). More importantly, no adverse reaction or rejection was observed although human MSCs and CM were used in a xenogeneic model. The presence of hFGF, hHGF, hGCSF, hIL-1Ra, hVEGF, and hIL-6 in the secretome may elucidate the regenerative potential of the xeno-free cell-based and cell-free approaches which have translational value for advanced wound care. The results revealed the therapeutic potential of both the cell-based and cell-free approaches for wound healing.

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The oncometabolite R-2-hydroxyglutarate dysregulates the differentiation of human mesenchymal stromal cells via inducing DNA hypermethylation

BMC Cancer ◽

10.1186/s12885-020-07744-x ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Lizhen Liu ◽

Kaimin Hu ◽

Jingjing Feng ◽

Huafang Wang ◽

Shan Fu ◽

...

Keyword(s):

Osteogenic Differentiation ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Chondrogenic Differentiation ◽

Cell Counting ◽

Human Mscs ◽

Mrna Levels ◽

Dependent Manner ◽

Dna Hypermethylation ◽

Cartilaginous Tumors

Abstract Background Isocitrate dehydrogenase (IDH1/2) gene mutations are the most frequently observed mutations in cartilaginous tumors. The mutant IDH causes elevation in the levels of R-enantiomer of 2-hydroxylglutarate (R-2HG). Mesenchymal stromal cells (MSCs) are reasonable precursor cell candidates of cartilaginous tumors. This study aimed to investigate the effect of oncometabolite R-2HG on MSCs. Methods Human bone marrow MSCs treated with or without R-2HG at concentrations 0.1 to 1.5 mM were used for experiments. Cell Counting Kit-8 was used to detect the proliferation of MSCs. To determine the effects of R-2HG on MSC differentiation, cells were cultured in osteogenic, chondrogenic and adipogenic medium. Specific staining approaches were performed and differentiation-related genes were quantified. Furthermore, DNA methylation status was explored by Illumina array-based arrays. Real-time PCR was applied to examine the signaling component mRNAs involved in. Results R-2HG showed no influence on the proliferation of human MSCs. R-2HG blocked osteogenic differentiation, whereas promoted adipogenic differentiation of MSCs in a dose-dependent manner. R-2HG inhibited chondrogenic differentiation of MSCs, but increased the expression of genes related to chondrocyte hypertrophy in a lower concentration (1.0 mM). Moreover, R-2HG induced a pronounced DNA hypermethylation state of MSC. R-2HG also improved promotor methylation of lineage-specific genes during osteogenic and chondrogenic differentiation. In addition, R-2HG induced hypermethylation and decreased the mRNA levels of SHH, GLI1and GLI2, indicating Sonic Hedgehog (Shh) signaling inhibition. Conclusions The oncometabolite R-2HG dysregulated the chondrogenic and osteogenic differentiation of MSCs possibly via induction of DNA hypermethylation, improving the role of R-2HG in cartilaginous tumor development.

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Comparative Study of Ceramics Structure for Culturing Human Mesenchymal Stromal Cells

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.361-363.1067 ◽

2007 ◽

Vol 361-363 ◽

pp. 1067-1070 ◽

Cited By ~ 1

Author(s):

Asako Matsushima ◽

Noriko Kotobuki ◽

Mika Tadokoro ◽

Hajime Ohgushi

Keyword(s):

Bone Tissue ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Cell Attachment ◽

Osteoblastic Differentiation ◽

Human Mscs ◽

Osteogenic Cells ◽

Human Mesenchymal Stromal Cells ◽

Viable Cells ◽

Different Types

Hydroxyapatite (HA) ceramics together with various kinds of osteogenic cells have been used in bone tissue engineering. It is well known that the ceramics structure and composition affect cell proliferation / differentiation. In this study, three different types of HA ceramics were used to investigate initial cell attachment followed by osteoblastic differentiation of human mesenchymal stromal cells (MSCs). The results indicated that micro-pore affected the cell attachment and porosity (pore diameter and inter-pore connection) was the key to allow spacious distribution of the viable cells in the ceramics. This study also confirmed that surface pore areas of HA ceramics support the differentiation of human MSCs and thus the ceramics have the capability to regenerate damaged bone tissue.

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