scholarly journals Timing of induction of cardiomyocyte differentiation for in vitro cultured mesenchymal stem cells: a perspective for emergenciesThis article is one of a selection of papers from the NATO Advanced Research Workshop on Translational Knowledge for Heart Health (published in part 1 of a 2-part Special Issue).

2009 ◽  
Vol 87 (2) ◽  
pp. 143-150 ◽  
Author(s):  
Zeynep Tokcaer-Keskin ◽  
A. Ruchan Akar ◽  
Fatma Ayaloglu-Butun ◽  
Ece Terzioglu-Kara ◽  
Serkan Durdu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Troponin I ◽  
Cell Lineage ◽  
Specific Cell ◽  
Cardiomyocyte Differentiation ◽  
Time Period ◽  
Intracellular Ca ◽  
Colony Forming Capacity

Mesenchymal stem cells (MSCs) have the capacity to differentiate into osteoblasts, chondrocytes, adipocytes, myocytes, and cardiomyocytes. Several established methods are presently available for in vitro isolation of MSCs from bone marrow. However, the duration necessary to culture them can be a major handicap to cell-based therapies needed for such urgent cardiovascular conditions as acute myocardial infarction and acute hindlimb ischemia. The best timing of cardiomyocyte differentiation induction after MCS isolation and expansion is still an unresolved issue. Our goal was to investigate the possibility of obtaining functional cardiomyocytes from rat MSC within a shorter time period. We examined MSCs’ colony-forming capacity, CD90 and CD34 immunoreactivity during the 14 days of culturing. Cardiomyocyte differentiation was induced by 5-azacytidine. Immunohistochemic staining, together with intracellular Ca2+ measurement experiments, revealed that MSCs do not differentiate into any specific cell lineage but show the characteristics of MSCs on both the 9th and 14th days of the culture. To check the potential for differentiation into cardiomyocytes, experiments with caffeine application and depolarization with KCl were performed. The cells possessed some of the specific biochemical features of contracting cells, with slightly higher capacities on the 14th day. Cells from 9th and 14th days of the culture that were treated with 5-azacytidine had a higher expression of cardiac-specific markers such as troponin I, α-sarcomeric actin, and MEF2D compared with the control groups. This study illustrates that it is possible to get functional cardiomyocytes from in vitro MSC culture in a shorter time period than previously achieved. This reduction in time may provide emergency cases with access to cell-based therapies that may have previously been unavailable.

scholarly journals Polymeric sheet actuators with programmable bioinstructivity

2020 ◽  
Vol 117 (4) ◽  
pp. 1895-1901 ◽  
Author(s):  
Zijun Deng ◽  
Weiwei Wang ◽  
Xun Xu ◽  
Oliver E. C. Gould ◽  
Karl Kratz ◽  
...  
Keyword(s):  
Stem Cells ◽  
Shape Change ◽  
Cell Lineage ◽  
Fundamental Principle ◽  
Signaling Cascades ◽  
Specific Cell ◽  
Polymer Actuator ◽  
Intracellular Ca ◽  
Functional Biomaterials ◽  
Biochemical Signals

Stem cells are capable of sensing and processing environmental inputs, converting this information to output a specific cell lineage through signaling cascades. Despite the combinatorial nature of mechanical, thermal, and biochemical signals, these stimuli have typically been decoupled and applied independently, requiring continuous regulation by controlling units. We employ a programmable polymer actuator sheet to autonomously synchronize thermal and mechanical signals applied to mesenchymal stem cells (MSCs). Using a grid on its underside, the shape change of polymer sheet, as well as cell morphology, calcium (Ca2+) influx, and focal adhesion assembly, could be visualized and quantified. This paper gives compelling evidence that the temperature sensing and mechanosensing of MSCs are interconnected via intracellular Ca2+. Up-regulated Ca2+ levels lead to a remarkable alteration of histone H3K9 acetylation and activation of osteogenic related genes. The interplay of physical, thermal, and biochemical signaling was utilized to accelerate the cell differentiation toward osteogenic lineage. The approach of programmable bioinstructivity provides a fundamental principle for functional biomaterials exhibiting multifaceted stimuli on differentiation programs. Technological impact is expected in the tissue engineering of periosteum for treating bone defects.

Sertoli cell-mediated differentiation of bovine fetal mesenchymal stem cells into germ cell lineage using an in vitro co-culture system

2019 ◽  
Vol 130 ◽  
pp. 8-18
Author(s):  
M.N. Segunda ◽  
J. Bahamonde ◽  
I. Muñoz ◽  
S. Sepulveda ◽  
J. Cortez ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Germ Cell ◽  
Sertoli Cell ◽  
Culture System ◽  
Cell Lineage

scholarly journals Improved Efficiency of Cardiomyocyte-Like Cell Differentiation from Rat Adipose Tissue-Derived Mesenchymal Stem Cells with a Directed Differentiation Protocol

2019 ◽  
Vol 2019 ◽  
pp. 1-16 ◽  
Author(s):  
Blanca Rebeca Ibarra-Ibarra ◽  
Martha Franco ◽  
Araceli Paez ◽  
Elvira Varela López ◽  
Felipe Massó
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Cell Differentiation ◽  
Cardiac Differentiation ◽  
Cardiomyocyte Differentiation ◽  
Cell Based Therapy ◽  
Differentiation Protocol ◽  
Differentiation Efficiency

Cell-based therapy has become a resource for the treatment of cardiovascular diseases; however, there are some conundrums to achieve. In vitro cardiomyocyte generation could be a solution for scaling options in clinical applications. Variability on cardiac differentiation in previously reported studies from adipose tissue-derived mesenchymal stem cells (ASCs) and the lack of measuring of the cardiomyocyte differentiation efficiency motivate the present study. Here, we improved the ASC-derived cardiomyocyte-like cell differentiation efficiency with a directed cardiomyocyte differentiation protocol: BMP-4 + VEGF (days 0-4) followed by a methylcellulose-based medium with cytokines (IL-6 and IL-3) (days 5-21). Cultures treated with the directed cardiomyocyte differentiation protocol showed cardiac-like cells and “rosette-like structures” from day 7. The percentage of cardiac troponin T- (cTnT-) positive cells was evaluated by flow cytometry to assess the cardiomyocyte differentiation efficiency in a quantitative manner. ASCs treated with the directed cardiomyocyte differentiation protocol obtained a differentiation efficiency of up to 44.03% (39.96%±3.78) at day 15 without any enrichment step. Also, at day 21 we observed by immunofluorescence the positive expression of early, late, and cardiac maturation differentiation markers (Gata-4, cTnT, cardiac myosin heavy chain (MyH), and the sarcoplasmic/endoplasmic reticulum Ca2+ ATPase (SERCa2)) in cultures treated with the directed cardiomyocyte differentiation protocol. Unlike other protocols, the use of critical factors of embryonic cardiomyogenesis coupled with a methylcellulose-based medium containing previously reported cardiogenic cytokines (IL-6 and IL-3) seems to be favorable for in vitro cardiomyocyte generation. This novel efficient culture protocol makes ASC-derived cardiac differentiation more efficient. Further investigation is needed to identify an ASC-derived cardiomyocyte surface marker for cardiac enrichment.

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

2019 ◽  
Vol 15 (6) ◽  
pp. 814-826 ◽  
Author(s):  
Sajan George ◽  
Michael R. Hamblin ◽  
Heidi Abrahamse
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adult Stem Cells ◽  
Molecular Mechanisms ◽  
Culture Media ◽  
Cell Signalling ◽  
Specific Cell ◽  
Ongoing Research ◽  
Neural Lineage

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.

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

2017 ◽  
Vol 5 (10) ◽  
pp. 2056-2067 ◽  
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.

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

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.

Abstract 192: Histological and Functional Assessment of Human Placental Stem Cells in an Induced Myocardial Infarction in Rats

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Georges Makhoul ◽  
Yu-Ting Ma ◽  
Minh Duong ◽  
Ray C J Chiu ◽  
Renzo Cecere
Keyword(s):  
Myocardial Infarction ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Troponin I ◽  
Cardiac Cells ◽  
Coronary Artery Ligation ◽  
Regenerative Therapy ◽  
Age Related ◽  
Group 1

Purpose: Human bone marrow mesenchymal stem cells (hbmMSCs) have been studied extensively for myocardial regenerative therapy. However, such cells require invasive procreation and suffer from donor age-related declining quality. Recently, a more abundant resource of young MSCs has been isolated from an otherwise discarded organ: the human placenta mesenchymal stem cells (hpMSCs). In this study, we wanted to examine the survival, differentiation, and functionality of xenogeneic hpMSCs when implanted into an induced myocardial rat infarction. Methods: To inspect their stemness,hpMSCs underwent an In Vitro cardiac cell differentiation in a DMEM medium containing 5 Azacytidine. Additionally, hpMSCs were tested in a myocardial infarction animal model. Female Lewis rats (40 animals) underwent left coronary artery ligation. Animals were divided into 4 groups. Group 1 was injected with hpMSCs in the peri-infarct region. Groups 2 and 3 received hbmMSCs and In Vitro differentiated hbmMSCs into cardiomyocytes respectively. Cell free medium was injected in group 4. Echocardiography was performed at baseline, day 4, weeks 3, 6, and 9 after ligation. Myocardial tissues were harvested and studied immunohistochemically for specific muscular and cardiac markers (Actin and Troponin I) on weeks 6 and 10. Results: In Vitro differentiation into cardiomyocyte lineage was achieved with the hpMSCs. HpMSCs were detected within rat myocardium by week 6 after their implantation. These cells stained positively for Actin and Troponin I. Preliminary echocardiographic data show cardiac functional increase in group 1. Whether hpMSCs can provide a superior effect than hbmMSCs or differentiated hbmMSCs is being investigated. Conclusions: In Vitro studies indicated that the hpMSCs can be differentiated into cardiac cells. When implanted into the rat infarcted myocardium, hpMSCs survived and were able to differentiate into cardiomyocytes. These cells appeared to increase cardiac function in an induced myocardial infarction rat model. With the advantages of easy availability and young age, hpMSCs could be more suitable for myocardial regenerative therapy.

scholarly journals Differentiation of Human Dermal Mesenchymal Stem Cells into Cardiomyocytes by Treatment with 5-Azacytidine: Concept for Regenerative Therapy in Myocardial Infarction

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Pravin D. Potdar ◽  
Preeti Prasannan
Keyword(s):  
Myocardial Infarction ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Troponin T ◽  
Punch Biopsy ◽  
Cardiomyocyte Differentiation ◽  
Mesenchymal Phenotype ◽  
Degenerative Disorders ◽  
Cardiac Therapy

Myocardial infarction (MI) is the leading cause of death worldwide. Stem cells regenerative medicine offers a promising approach to cure such degenerative disorders. Mesenchymal stem cells are thought to be one of the important types of stem cells which can differentiate into various lineages such as neuron, hepatocytes, and cardiomyocytes. In the present study, human dermal mesenchymal stem cells (hDMSCs) have been developed from human scalp punch biopsy and characterized for their mesenchymal phenotype so that these cells can be useful for differentiating into cardiomyocytes. 5-Azacytidine induces cardiomyocyte differentiation in vitro and therefore it has been used to differentiate hDMSCs cells into cardiomyocytes. It was observed that hDMSCs differentiated into cardiomyocyte within a period of 4 days to 15 days after treatment with 10 μM and 20 μM of 5-azacytidine. The cardiomyocyte phenotype was confirmed by studying expression of α-cardiac actin, β-myosin heavy chain, and cardiac troponin T. Thus, this paper describes the differentiation of hDMSCs into cardiomyocytes which can be further be used for treatment of MI. This type of cell-based cardiac therapy will offer a new hope for millions of patients worldwide who are suffering from heart disease.

scholarly journals Reorganization of Metabolism during Cardiomyogenesis Implies Time-Specific Signaling Pathway Regulation

2021 ◽  
Vol 22 (3) ◽  
pp. 1330
Author(s):  
María Julia Barisón ◽  
Isabela Tiemy Pereira ◽  
Anny Waloski Robert ◽  
Bruno Dallagiovanna
Keyword(s):  
Stem Cells ◽  
Embryoid Body ◽  
Embryonic Stem ◽  
Cell Lineage ◽  
Cardiac Differentiation ◽  
Lipid Homeostasis ◽  
Specific Cell ◽  
Regulatory Pathways ◽  
Time Specific

Understanding the cell differentiation process involves the characterization of signaling and regulatory pathways. The coordinated action involved in multilevel regulation determines the commitment of stem cells and their differentiation into a specific cell lineage. Cellular metabolism plays a relevant role in modulating the expression of genes, which act as sensors of the extra-and intracellular environment. In this work, we analyzed mRNAs associated with polysomes by focusing on the expression profile of metabolism-related genes during the cardiac differentiation of human embryonic stem cells (hESCs). We compared different time points during cardiac differentiation (pluripotency, embryoid body aggregation, cardiac mesoderm, cardiac progenitor and cardiomyocyte) and showed the immature cell profile of energy metabolism. Highly regulated canonical pathways are thoroughly discussed, such as those involved in metabolic signaling and lipid homeostasis. We reveal the critical relevance of retinoic X receptor (RXR) heterodimers in upstream retinoic acid metabolism and their relationship with thyroid hormone signaling. Additionally, we highlight the importance of lipid homeostasis and extracellular matrix component biosynthesis during cardiomyogenesis, providing new insights into how hESCs reorganize their metabolism during in vitro cardiac differentiation.

scholarly journals IFN-γ Licensing Does Not Enhance the Reduced Immunomodulatory Potential and Migratory Ability of Differentiation-Induced Porcine Bone Marrow-Derived Mesenchymal Stem Cells in an In Vitro Xenogeneic Application

2021 ◽  
Vol 2021 ◽  
pp. 1-15
Author(s):  
Hyeon-Jeong Lee ◽  
Hwan-Deuk Kim ◽  
Chan-Hee Jo ◽  
Eun-Yeong Bok ◽  
Saet-Byul Kim ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Mononuclear Cells ◽  
Therapeutic Potential ◽  
Specific Gene ◽  
Specific Cell ◽  
Ifn Γ

IFN-γ licensing to mesenchymal stem cells (MSCs) is applied to enhance the therapeutic potential of MSCs. However, although the features of MSCs are affected by several stimuli, little information is available on changes to the therapeutic potential of IFN-γ-licensed differentiated MSCs during xenogeneic applications. Therefore, the present study is aimed at clarifying the effects of adipogenic/osteogenic differentiation and IFN-γ licensing on the in vitro immunomodulatory and migratory properties of porcine bone marrow-derived MSCs in xenogeneic applications using human peripheral blood mononuclear cells (PBMCs). IFN-γ licensing in differentiated MSCs lowered lineage-specific gene expression but did not affect MSC-specific cell surface molecules. Although indoleamine 2,3 deoxygenase (IDO) activity and expression were increased after IFN-γ licensing in undifferentiated MSCs, they were reduced after differentiation. IFN-γ licensing to differentiated MSCs elevated the reduced IDO expression in differentiated MSCs; however, the increase was not sufficient to reach to the level achieved by undifferentiated MSCs. During a mixed lymphocyte reaction with quantification of TNF-α concentration, proliferation and activation of xenogeneic PBMCs were suppressed by undifferentiated MSCs but inhibited to a lesser extent by differentiated MSCs. IFN-γ licensing increasingly suppressed proliferation of PBMCs in undifferentiated MSCs but it was incapable of elevating the reduced immunosuppressive ability of differentiated MSCs. Migratory ability through a scratch assay and gene expression study was reduced in differentiated MSCs than their undifferentiated counterparts; IFN-γ licensing was unable to enhance the reduced migratory ability in differentiated MSCs. Similar results were found in a Transwell system with differentiated MSCs in the upper chamber toward xenogeneic PBMCs in the lower chamber, despite IFN-γ licensing increased the migratory ability of undifferentiated MSCs. Overall, IFN-γ licensing did not enhance the reduced immunomodulatory and migratory properties of differentiated MSCs in a xenogeneic application. This study provides a better understanding of the ways in which MSC therapy can be applied.

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