Differentiation of Mesenchymal Stem Cells to Neuroglia: in the Context of Cell Signalling

Abstract The promise of engineering specific cell types from stem cells and rebuilding damaged or diseased tissues has fascinated stem cell researchers and clinicians over last few decades. Mesenchymal Stem Cells (MSCs) have the potential to differentiate into non-mesodermal cells, particularly neural-lineage, consisting of neurons and glia. These multipotent adult stem cells can be used for implementing clinical trials in neural repair. Ongoing research identifies several molecular mechanisms involved in the speciation of neuroglia, which are tightly regulated and interconnected by various components of cell signalling machinery. Growing MSCs with multiple inducers in culture media will initiate changes on intricately interlinked cell signalling pathways and processes. Net result of these signal flow on cellular architecture is also dependent on the type of ligands and stem cells investigated in vitro. However, our understanding about this dynamic signalling machinery is limited and confounding, especially with spheroid structures, neurospheres and organoids. Therefore, the results for differentiating neurons and glia in vitro have been inconclusive, so far. Added to this complication, we have no convincing evidence about the electrical conductivity and functionality status generated in differentiating neurons and glia. This review has taken a step forward to tailor the information on differentiating neuroglia with the common methodologies, in practice.

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Mesenchymal Stem Cells in the Treatment of COVID-19, a Promising Future

Cells ◽

10.3390/cells10102588 ◽

2021 ◽

Vol 10 (10) ◽

pp. 2588

Author(s):

Daniela Gois Beghini ◽

Samuel Iwao Horita ◽

Andrea Henriques-Pons

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Intravenous Injection ◽

Adult Stem Cells ◽

Ethical Issues ◽

Genetic Manipulation ◽

Culture Media ◽

Embryonic Stem

Mesenchymal stem cells (MSCs) are multipotent adult stem cells present in virtually all tissues; they have a potent self-renewal capacity and can differentiate into multiple cell types. They also affect the ambient tissue by the paracrine secretion of numerous factors in vivo, including the induction of other stem cells’ differentiation. In vitro, the culture media supernatant is named secretome and contains soluble molecules and extracellular vesicles that retain potent biological function in tissue regeneration. MSCs are considered safe for human treatment; their use does not involve ethical issues, as embryonic stem cells do not require genetic manipulation as induced pluripotent stem cells, and after intravenous injection, they are mainly found in the lugs. Therefore, these cells are currently being tested in various preclinical and clinical trials for several diseases, including COVID-19. Several affected COVID-19 patients develop induced acute respiratory distress syndrome (ARDS) associated with an uncontrolled inflammatory response. This condition causes extensive damage to the lungs and may leave serious post-COVID-19 sequelae. As the disease may cause systemic alterations, such as thromboembolism and compromised renal and cardiac function, the intravenous injection of MSCs may be a therapeutic alternative against multiple pathological manifestations. In this work, we reviewed the literature about MSCs biology, focusing on their function in pulmonary regeneration and their use in COVID-19 treatment.

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Melatonin Reverses the Loss of Stemness Induced by TNF-α in Human Bone Marrow Mesenchymal Stem Cells through Upregulation of YAP Expression

Stem Cells International ◽

10.1155/2019/6568394 ◽

2019 ◽

Vol 2019 ◽

pp. 1-16 ◽

Cited By ~ 4

Author(s):

Xudong Wang ◽

Tongzhou Liang ◽

Jincheng Qiu ◽

Xianjian Qiu ◽

Bo Gao ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Molecular Mechanisms ◽

Inflammatory Factor ◽

Cell Transplant ◽

Alizarin Red ◽

Marrow Mesenchymal Stem Cells

Mesenchymal stem cells (MSCs) are promising candidates for tissue regeneration and disease treatment. However, long-term in vitro culture results in loss of MSC stemness. The inflammation that occurs at stem cell transplant sites (such as that resulting from TNF-α) is a contributing factor for stem cell treatment failure. Currently, there is little evidence regarding the protective role of melatonin with regard to the negative effects of TNF-α on the stemness of MSCs. In this study, we report a melatonin-based method to reduce the inflammatory effects on the stemness of bone marrow mesenchymal stem cells (BMMSCs). The results of colony formation assays, Alizarin red staining, western blotting, and reverse transcription-polymerase chain reactions suggest that melatonin can reverse the inflammatory damage caused by TNF-α treatment in the third, seventh, and tenth generations of primary BMMSCs (vs. control and the TNF-α-treated group). Meanwhile, a detailed analysis of the molecular mechanisms showed that the melatonin receptor and YAP signaling pathway are closely related to the role that melatonin plays in negative inflammatory effects against BMMSCs. In addition, in vivo experiments showed that melatonin could reverse the damage caused by TNF-α on bone regeneration by BMMSCs in nude mice. Overall, our results suggest that melatonin can reverse the loss of stemness caused by inflammatory factor TNF-α in BMMSCs. Our results also provide a practical strategy for the application of BMMSCs in tissue engineering and cell therapy.

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Environmental Benzopyrene Attenuates Stemness of Placenta-Derived Mesenchymal Stem Cells via Aryl Hydrocarbon Receptor

Stem Cells International ◽

10.1155/2019/7414015 ◽

2019 ◽

Vol 2019 ◽

pp. 1-12 ◽

Cited By ~ 4

Author(s):

June Seok Heo ◽

Ja-Yun Lim ◽

Sangshin Pyo ◽

Dae Wui Yoon ◽

Dongsook Lee ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Aryl Hydrocarbon Receptor ◽

Molecular Mechanisms ◽

Cell Activity ◽

Bone Diseases ◽

Purine Derivative ◽

Aryl Hydrocarbon ◽

Polycyclic Aromatic

The toxic effects of particulate matter have been linked to polycyclic aromatic hydrocarbons (PAHs) such as benzopyrene. PAHs are potent inducers of the aryl hydrocarbon receptor (AhR), which is an expressed nuclear receptor that senses environmental stimuli and modulates gene expression. Even though several studies have shown that the benzopyrene (BP) of chemical pollutants significantly impaired stem cell activity, the exact molecular mechanisms were not clearly elucidated. In the present study, we aimed to investigate the effects of BP on placenta-derived mesenchymal stem cells (PD-MSCs) in vitro. We found that the AhR in PD-MSCs was expressed under the treatment of BP, and its activation markedly disrupted osteogenic differentiation through the alteration of stemness activity of PD-MSCs. Moreover, BP treatment significantly reduced the proliferation activity of PD-MSCs and expression of pluripotent markers through the induction of AhR. Treatment with StemRegenin 1 (SR1), a purine derivative that antagonizes the AhR, effectively prevented BP-induced reduction of the proliferation and differentiation activity of PD-MSCs. In this study, we found that BP treatment in PD-MSCs markedly obstructs PD-MSC stemness through AhR signaling. Noteworthy, SR1-mediated MSC application will contribute to new perspectives on MSC-based therapies for air pollution-related bone diseases.

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MMP-2, MT1-MMP, and TIMP-2 are essential for the invasive capacity of human mesenchymal stem cells: differential regulation by inflammatory cytokines

Blood ◽

10.1182/blood-2006-10-051060 ◽

2007 ◽

Vol 109 (9) ◽

pp. 4055-4063 ◽

Cited By ~ 331

Author(s):

Christian Ries ◽

Virginia Egea ◽

Marisa Karow ◽

Helmut Kolb ◽

Marianne Jochum ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Migration ◽

Inflammatory Cytokines ◽

Molecular Mechanisms ◽

Human Mesenchymal Stem Cells ◽

Basement Membranes ◽

Tissue Inhibitor Of Metalloproteinase ◽

Matrix Metalloproteinase 2

Abstract Human mesenchymal stem cells (hMSCs) represent promising tools in various clinical applications, including the regeneration of injured tissues by endogenous or transplanted hMSCs. The molecular mechanisms, however, that control hMSC mobilization and homing which require invasion through extracellular matrix (ECM) barriers are almost unknown. We have analyzed bone marrow–derivedhMSCs and detected strong expression and synthesis of matrix metalloproteinase 2 (MMP-2), membrane type 1 MMP (MT1-MMP), tissue inhibitor of metalloproteinase 1 (TIMP-1), and TIMP-2. The ability of hMSCs to traverse reconstituted human basement membranes was effectively blocked in the presence of synthetic MMP inhibitors. Detailed studies by RNA interference revealed that gene knock-down of MMP-2, MT1-MMP, or TIMP-2 substantially impaired hMSC invasion, whereas silencing of TIMP-1 enhanced cell migration, indicating opposing roles of both TIMPs in this process. Moreover, the inflammatory cytokines TGF-β1, IL-1β, and TNF-α up-regulated MMP-2, MT1-MMP, and/or MMP-9 production in these cells, resulting in a strong stimulation of chemotactic migration through ECM, whereas the chemokine SDF-1α exhibited minor effects on MMP/TIMP expression and cell invasion. Thus, induction of specific MMP activity in hMSCs by inflammatory cytokines promotes directed cell migration across reconstituted basement membranes in vitro providing a potential mechanism in hMSC recruitment and extravasation into injured tissues in vivo.

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Combined effects of multi-scale topographical cues on stable cell sheet formation and differentiation of mesenchymal stem cells

Biomaterials Science ◽

10.1039/c7bm00134g ◽

2017 ◽

Vol 5 (10) ◽

pp. 2056-2067 ◽

Cited By ~ 6

Author(s):

Sisi Li ◽

Shreyas Kuddannaya ◽

Yon Jin Chuah ◽

Jingnan Bao ◽

Yilei Zhang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Sheet ◽

Specific Cell ◽

Combined Effects ◽

Multi Scale ◽

Cell Responses ◽

Topographical Cues

To decipher specific cell responses to diverse and complex in vivo signals, it is essential to emulate specific surface chemicals, extra cellular matrix (ECM) components and topographical signals through reliable and easily reproducible in vitro systems.

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Behavior of Mesenchymal Stem Cells Obtained From Dental Tissues: A Review of the Literature

Odovtos - International Journal of Dental Sciences ◽

10.15517/ijds.v21i1.34884 ◽

2019 ◽

Vol 21 (1) ◽

pp. 31-40

Author(s):

Mariné Ortiz-Magdaleno DDS, MSc, PhD ◽

Ana Isabel Romo-Tobías DDS ◽

Fernando Romo-Ramírez DDS, MSc ◽

Diana María Escobar DDS, MSc, PhD ◽

Héctor Flores-Reyes DDS, MSc, PhD ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Clinical Evidence ◽

Periodontal Regeneration ◽

In Vivo Studies ◽

Specific Cell ◽

Key Factors ◽

Dental Tissues

The success of tissue engineering in combination with tissue regeneration depends on the behavior and cellular activity in the biological processes developed within a structure that functions as a support, better known as scaffolds, or directly at the site of the injury. The cell-cell and cell-biomaterial interaction are key factors for the induction of a specific cell behavior, together with the bioactive factors that allow the formation of the desired tissue. Mesenchymal Stem Cells (MSC) can be isolated from the umbilical cord and bone marrow; however, the behavior of Dental Pulp Stem Cells (DPSC) has been shown to have a high potential for the formation of bone tissue, and these cells have even been able to induce the process of angiogenesis. Advances in periodontal regeneration, dentin-pulp complex, and craniofacial bone defects through the induction of MSC obtained from tooth structures in in vitro-in vivo studies have permitted the obtaining of clinical evidence of the achievements obtained to date.

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Effect of conditioned medium of umbilical cord-derived mesenchymal stem cells as a culture medium for human granulosa cells: An experimental study

International Journal of Reproductive BioMedicine ◽

10.18502/ijrm.v19i12.10054 ◽

2022 ◽

pp. 1037-1044

Author(s):

Kanadi Sumapraja ◽

Andon Hestiantoro ◽

Isabella Kurnia Liem ◽

Arief Boediono ◽

Teuku Z Jacoeb

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Conditioned Medium ◽

Umbilical Cord ◽

Granulosa Cells ◽

Culture Medium ◽

Culture Media ◽

Controlled Ovarian Hyperstimulation ◽

Vitro Fertilization

Background: The umbilical cord-derived mesenchymal stem cells conditioned medium (UC-MSCs-CM) produces secretomes with anti-apoptotic properties, and has the potential to prevent apoptosis of granulosa cells (GC) during controlled ovarian hyperstimulation. Objective: To observe the effect of UC-MSCs-CM on the interaction between pro- and anti-apoptotic proteins and the influence of growth differentiation factor 9 (GDF9) production in GC. Materials and Methods: UC-MSCs-CM was collected from umbilical cord stem cell culture on passage 4. GC from 23 women who underwent in vitro fertilization were cultured and exposed to UC-MSCs-CM for 24 hr. Then RNA of the GC was extracted and the mRNA expression of BCL-2 associated X (BAX), survivin and GDF9 were analysed using quantitative real-time PCR. The spent culture media of the GC were collected for measurement of insulin growth factor 1 using ELISA. Results: The expression of BAX was significantly different after UC-MSCs-CM exposure (4.09E-7 vs. 3.74E-7, p = 0.02). No significant changes occurred in survivin, BAX/survivin ratio, and GDF9 expression after UC-MSCs-CM exposure (p > 0.05). The IGF-1 level of the CM was significantly higher after the CM was used as a culture medium for GC (2.28 vs. 3.07 ± 1.72, p ≤ 0.001). A significant positive correlation was found between survivin and GDF9 (r = 0.966, p ≤ 0.001). Conclusion: IGF-1 produced by UC-MSCs-CM can work in paracrine fashion through the IGF receptor, which can inhibit BAX and maintain GDF9 production. Moreover, under the influence of UC-MSCs-CM, GC are also capable of producing IGF-1, which can impact GC through autocrine processes. Key words: Conditioned medium, BAX, Survivin, GDF9, IGF-1.

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213 TRANSCRIPTOMIC COMPARISON BETWEEN PORCINE ADIPOSE AND BONE MARROW MESENCHYMAL STEM CELLS DURING IN VITRO OSTEOGENIC AND ADIPOGENIC DIFFERENTIATION

Reproduction Fertility and Development ◽

10.1071/rdv24n1ab213 ◽

2012 ◽

Vol 24 (1) ◽

pp. 219

Author(s):

E. Monaco ◽

M. Bionaz ◽

A. Lima ◽

W. L. Hurley ◽

M. B. Wheeler

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Lipid Metabolism ◽

Adult Stem Cells ◽

Specific Factor ◽

Specific Gene ◽

Fatty Acid Binding ◽

Gene Markers

Previous data support the use of adipose-derived stem cells (ASC) as an alternative to bone marrow as a source of adult stem cells for therapeutic purposes. The aims of the present study were to directly compare the transcriptome of ASC and bone marrow-derived mesenchymal stem cells (BMSC) in order to uncover potential functional differences between the two mesenchymal stem cells (MSC), to identify differentially expressed genes (DEG) and related functions that may drive MSC to become bone or adipose and to identify potential markers for adipogenic and osteogenic differentiation. ASC and BMSC, isolated from subcutaneous adipose tissue and femurs of 3 adult pigs were differentiated in vitro along the osteogenic and adipogenic lineage for up to 4 wk. At 0, 2, 7 and 21 days of differentiation, RNA was extracted for microarray analysis. Data were normalized by Lowess and statistical analysis run using ANOVA with Benjamini-Hochberg false discovery rate (FDR) correction. Data mining was carried out using Ingenuity Pathway Analysis and DAVID. Analysis of undifferentiated MSC for genes with the highest expression and DEG between MSC and fully differentiated tissues uncovered MSC being featured by low immunity and high angiogenic capacity. The direct comparison between differentiation lineages indicated that the expression of a limited number of genes has to change in order to determine cell fate. Functional analysis of the DEG between differentiation lineages indicated that osteogenesis is characterised by larger cell proliferation and cytoskeleton organisation with a crucial role of G-proteins compared to adipogenesis. On the other hand, adipogenesis is driven by PPAR signalling, has greater angiogenesis, lipid metabolism, migration and tumorigenesis capacity compared to osteogenesis. The direct comparison between ASC and BMSC during the same differentiation uncovered that ASC is featured by a greater lipid metabolism compared to BMSC, while BMSC has a more pronounced cell growth and proliferation than ASC. In addition, we uncovered 39 specific gene markers for adipogenesis and 65 for osteogenesis. NAD(P)H dehydrogenase quinone 1 (NQO1), aquaporin 3 (AQP3), stearoyl-CoA desaturase (SCD), fatty acid binding protein 3 and 5 (FABP3 and FABP5) and ferritin light polypeptide (FTL) were among the best adipogenic markers. Hemopexin (HPX), collagenase type 3α (COL3A1), annexin A8-like 1 (ANXA8L1), flotillin 2 (FLOT2) and periostin or osteoblast specific factor (POSTN) were among the best osteogenic markers. Overall, the data indicated that the transcriptome of the two MSC are similar across the conditions studied. In addition, despite the limited DEG between the two MSC, the enrichment of several functions/pathways might indicate differences in therapeutic application. This work was support by the Illinois Regenerative Medicine Institute (IDPH # 63080017).

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Abstract 515: Non-invasive Method of Generating Human Induced-mesenchymal Stem Cells Derived From Urinary Epithelial Cells for Regenerative Therapy

Circulation Research ◽

10.1161/res.127.suppl_1.515 ◽

2020 ◽

Vol 127 (Suppl_1) ◽

Author(s):

Narasimman Gurusamy ◽

SHEEJA RAJASINGH ◽

Vijay Selvam ◽

Vinoth Sigamani ◽

Jayavardini Vasanthan ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Epithelial Cells ◽

Adult Stem Cells ◽

Regenerative Therapy ◽

Non Invasive ◽

Proliferation Capacity ◽

Invasive Method

Introduction: Mesenchymal stem cells (MSCs) are multipotent adult stem cells having an extensive proliferation capacity in vitro and in vivo. These MSCs can differentiate into various mesoderm-type cells such as osteoblasts, cardiomyocytes, etc. A subpopulation of urinary epithelial cells (UECs) have been identified in urine samples, is considered a promising cell resource for generating autologous induced-pluripotent stem cells (iPSCs). Hypothesis: We hypothesize that the production of high quality, autologous, induced-MSCs (iMSCs) with high replicative potential suitable for the regenerative therapy, using an easy, and the most non-invasive method of isolation, from human UECs. Methods and Results: Human urine was collected and centrifuged to obtain the UECs, which were characterized by the expression of CK19 and ZO1. These UECs were reprogrammed to iPSCs using a cocktail of mRNAs (OCT4, KLF4, SOX2, c-MYC, Nanog and Lin28) along with Lipofectamine for 11 days in culture. These iPSCs were characterized by the expression of the pluripotent markers such as OCT4, SOX2 and SSEA4. The iPSCs were subsequently differentiated into iMSCs using the mesenchymal specific medium for 21 days. iMSCs were harvested at the end of 21 days, and they were characterized by the high levels of mRNA and protein expressions of mesenchymal specific markers such as CD73, CD90 and CD105 (Fig. 1A). FACS analysis showed that more than 93% of the cells were positive for the markers of MSCs (Fig. 1B) . Moreover, the obtained iMSCs have high proliferation capacity compared with the adult stem cells. Conclusions: We have developed an easy, non-invasive method for obtaining autologous, non-immunogenic and highly-proliferating iMSCs suitable for various regenerative therapies including cardiac diseases, from urinary epithelial cells.

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Rapid Rapamycin-Only Induced Osteogenic Differentiation of Blood-Derived Stem Cells and Their Adhesion to Natural and Artificial Scaffolds

Stem Cells International ◽

10.1155/2017/2976541 ◽

2017 ◽

Vol 2017 ◽

pp. 1-12

Author(s):

Carpentieri Arianna ◽

Cozzoli Eliana ◽

Acri Flavio ◽

Ranalli Marco ◽

Diedenhofen Giacomo ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Stem Cell ◽

Osteogenic Differentiation ◽

Molecular Mechanisms ◽

Maxillofacial Surgery ◽

Ongoing Research ◽

Medicine Research ◽

Cell Based Therapy

Stem cells are a centerpiece of regenerative medicine research, and the recent development of adult stem cell-based therapy systems has vigorously expanded the scope and depth of this scientific field. The regeneration of damaged and/or degraded bone tissue in orthopedic, dental, or maxillofacial surgery is one of the main areas where stem cells and their regenerative potential could be used successfully, requiring tissue engineering solutions incorporating an ideal stem cell type paired with the correct mechanical support. Our contribution to this ongoing research provides a new model of in vitro osteogenic differentiation using blood-derived stem cells (BDSCs) and rapamycin, visibly expressing typical osteogenic markers within ten days of treatment. In depth imaging studies allowed us to observe the adhesion, proliferation, and differentiation of BDSCs to both titanium and bone scaffolds. We demonstrate that BDSCs can differentiate towards the osteogenic lineage rapidly, while readily adhering to the scaffolds we exposed them to. Our results show that our model can be a valid tool to study the molecular mechanisms of osteogenesis while tailoring tissue engineering solutions to these new insights.

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