Modulation of Human Mesenchymal Stem Cells by Electrical Stimulation Using an Enzymatic Biofuel Cell

Enzymatic biofuel cells (EBFCs) have excellent potential as components in bioelectronic devices, especially as active biointerfaces to regulate stem cell behavior for regenerative medicine applications. However, it remains unclear to what extent EBFC-generated electrical stimulation can regulate the functional behavior of human adipose-derived mesenchymal stem cells (hAD-MSCs) at the morphological and gene expression levels. Herein, we investigated the effect of EBFC-generated electrical stimulation on hAD-MSC cell morphology and gene expression using next-generation RNA sequencing. We tested three different electrical currents, 127 ± 9, 248 ± 15, and 598 ± 75 nA/cm2, in mesenchymal stem cells. We performed transcriptome profiling to analyze the impact of EBFC-derived electrical current on gene expression using next generation sequencing (NGS). We also observed changes in cytoskeleton arrangement and analyzed gene expression that depends on the electrical stimulation. The electrical stimulation of EBFC changes cell morphology through cytoskeleton re-arrangement. In particular, the results of whole transcriptome NGS showed that specific gene clusters were up- or down-regulated depending on the magnitude of applied electrical current of EBFC. In conclusion, this study demonstrates that EBFC-generated electrical stimulation can influence the morphological and gene expression properties of stem cells; such capabilities can be useful for regenerative medicine applications such as bioelectronic devices.

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Modulation of human mesenchymal stem cells by electrical stimulation using an enzymatic biofuel cell

10.21203/rs.3.rs-78184/v1 ◽

2020 ◽

Author(s):

Won-Yong Jeon ◽

Seyoung Mun ◽

WeiBeng Ng ◽

Keunsoo Kang ◽

Kyudong Han ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Electrical Stimulation ◽

Regenerative Medicine ◽

Cell Morphology ◽

Wearable Sensors ◽

Electrical Current ◽

Next Generation ◽

The Impact

Abstract Background Enzymatic biofuel cells (EBFCs) have excellent potential as components in wearable sensors and bioelectronic devices, especially as active biointerfaces to regulate stem cell behavior for regenerative medicine applications. However, it remains unclear to what extent EBFC-generated electrical stimulation can regulate the functional behavior of human adipose-derived mesenchymal stem cells (hAD-MSCs) at the morphological and gene expression levels. Herein, we investigated the effect of EBFC-generated electrical stimulation on hAD-MSC cell morphology and gene expression using next-generation RNA sequencing. Materials We tested three different electrical currents,127 ± 9, 248 ± 15, and 598 ± 75 nA/cm2, in mesenchymal stem cells. We performed transcriptome profiling to analyze the impact of EBFC-derived electrical current on gene expression using next generation sequencing (NGS). Also we observed changes in cytoskeleton arrangement and analyzed gene expression depends on the electrical stimulation. Results The electrical stimulation of EBFC changes cell morphology through cytoskeleton re-arrangement. In particular, the results of whole transcriptome NGS showed that specific gene clusters were up- or down-regulated depending on the magnitude of applied electrical current of EBFC. Conclusion Taken together, the findings in this study demonstrate that EBFC-generated electrical stimulation can influence the morphological and gene expression properties of stem cells and such capabilities can be useful for regenerative medicine applications related to wearable sensors and devices.

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Temporal and Spatial Changes in Cartilage-Matrix-Specific Gene Expression in Mesenchymal Stem Cells in Response to Dynamic Compression

Tissue Engineering Part A ◽

10.1089/ten.tea.2011.0198 ◽

2011 ◽

Vol 17 (23-24) ◽

pp. 3085-3093 ◽

Cited By ~ 33

Author(s):

Matthew G. Haugh ◽

Eric G. Meyer ◽

Stephen D. Thorpe ◽

Tatiana Vinardell ◽

Garry P. Duffy ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Dynamic Compression ◽

Cartilage Matrix ◽

Specific Gene ◽

Specific Gene Expression ◽

Spatial Changes ◽

Temporal And Spatial

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Correlation between cell morphology and aggrecan gene expression level during differentiation from mesenchymal stem cells to chondrocytes

Biotechnology Letters ◽

10.1007/s10529-008-9683-8 ◽

2008 ◽

Vol 30 (7) ◽

pp. 1189-1195 ◽

Cited By ~ 19

Author(s):

Mutsumi Takagi ◽

Takayuki Kitabayashi ◽

Satoru Koizumi ◽

Haruka Hirose ◽

Shin-ichi Kondo ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Morphology ◽

Gene Expression Level ◽

Expression Level

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Effect of Zinc Ions on Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells to Male Germ Cells and Some Germ Cell-Specific Gene Expression in Rams

Biological Trace Element Research ◽

10.1007/s12011-012-9484-8 ◽

2012 ◽

Vol 150 (1-3) ◽

pp. 137-146 ◽

Cited By ~ 19

Author(s):

Mohammad Ghasemzadeh-Hasankolai ◽

Roozali Batavani ◽

Mohamadreza Baghaban Eslaminejad ◽

Mohammadali Sedighi-Gilani

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Germ Cell ◽

Germ Cells ◽

Specific Gene ◽

Zinc Ions ◽

Male Germ Cells ◽

Specific Gene Expression

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Comparative Analysis of Tenogenic Gene Expression in Tenocyte-Derived Induced Pluripotent Stem Cells and Bone Marrow-Derived Mesenchymal Stem Cells in Response to Biochemical and Biomechanical Stimuli

Stem Cells International ◽

10.1155/2021/8835576 ◽

2021 ◽

Vol 2021 ◽

pp. 1-18

Author(s):

Feikun Yang ◽

Dean W. Richardson

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Ectopic Expression ◽

Mechanical Stretch ◽

Specific Gene ◽

Induced Pluripotent

The tendon is highly prone to injury, overuse, or age-related degeneration in both humans and horses. Natural healing of injured tendon is poor, and cell-based therapeutic treatment is still a significant clinical challenge. In this study, we extensively investigated the expression of tenogenic genes in equine bone marrow mesenchymal stem cells (BMSCs) and tenocyte-derived induced pluripotent stem cells (teno-iPSCs) stimulated by growth factors (TGF-β3 and BMP12) combined with ectopic expression of tenogenic transcription factor MKX or cyclic uniaxial mechanical stretch. Western blotting revealed that TGF-β3 and BMP12 increased the expression of transcription factors SCX and MKX in both cells, but the tenocyte marker tenomodulin (TNMD) was detected only in BMSCs and upregulated by either inducer. On the other hand, quantitative real-time PCR showed that TGF-β3 increased the expression of EGR1, COL1A2, FMOD, and TNC in BMSCs and SCX, COL1A2, DCN, FMOD, and TNC in teno-iPSCs. BMP12 treatment elevated SCX, MKX, DCN, FMOD, and TNC in teno-iPSCs. Overexpression of MKX increased SCX, DCN, FMOD, and TNC in BMSCs and EGR1, COL1A2, DCN, FMOD, and TNC in teno-iPSCs; TGF-β3 further enhanced TNC in BMSCs. Moreover, mechanical stretch increased SCX, EGR1, DCN, ELN, and TNC in BMSCs and SCX, MKX, EGR1, COL1A2, DCN, FMOD, and TNC in teno-iPSCs; TGF-β3 tended to further elevate SCX, ELN, and TNC in BMSCs and SCX, MKX, COL1A2, DCN, and TNC in teno-iPSCs, while BMP12 further uptrended the expression of SCX and DCN in BMSCs and DCN in teno-iPSCs. Additionally, the aforementioned tenogenic inducers also affected the expression of signaling regulators SMAD7, ETV4, and SIRT1 in BMSCs and teno-iPSCs. Taken together, our data demonstrate that, in respect to the tenocyte-lineage-specific gene expression, BMSCs and teno-iPSCs respond differently to the tenogenic stimuli, which may affect the outcome of their application in tendon repair or regeneration.

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Improvement of in situ Follicular Activation and Early Development in Cryopreserved Human Ovarian Cortical Tissue by Co-Culturing with Mesenchymal Stem Cells

Cells Tissues Organs ◽

10.1159/000506303 ◽

2019 ◽

Vol 208 (1-2) ◽

pp. 48-58

Author(s):

Marzieh Hosseini ◽

Saghar Salehpour ◽

Marefat Ghaffari Novin ◽

Zahra Shams Mofarahe ◽

Mohammad-Amin Abdollahifar ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Cell Proliferation ◽

Tissue Culture ◽

Mesenchymal Stem Cells ◽

Ovarian Tissue ◽

Follicular Development ◽

Cortical Tissue ◽

Culture Systems ◽

The Impact

Follicular loss and tissue degeneration are great challenges in ovarian tissue culture systems. Mesenchymal stem cells (MSC) secrete a cocktail of growth factors and cytokines which supports adjacent cells and tissues. The aim of the current study was to investigate the impact of human bone marrow (hBM)-MSC, as co-culture cells, on human follicular development in ovarian cortical tissue (OCT) culture. For this purpose, warmed OCT fragments were co-cultured with hBM-MSC for 8 days and compared to monocultured OCT. During the culture period, ovarian follicle survival and development in the OCT were evaluated using histological observation, follicular developmental-related genes expression, and estradiol production. Furthermore, cell proliferation and apoptosis were assessed. The results showed that there were no significant differences in conserved ovarian follicles with a normal morphology between the two groups. However, the percentage of developing follicles, as well as follicular developmental gene expression, significantly increased in the co-culture group compared to the monoculture group. On the other hand, compared with the monoculture group, the co-culture group demonstrated a significant increase in cell proliferation, indicated by Ki67 gene expression, as well as a dramatic decrease in apoptotic cell percentage, revealed by TUNEL assay. These findings indicated that co-culturing of hBM-MSC with OCT could improve follicular activation and early follicular development in human ovarian tissue culture systems.

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128 Next-generation RNA sequencing of horse adipose and endometrial mesenchymal stem cells from the same donors unveils striking differences in their transcriptomic pattern

Reproduction Fertility and Development ◽

10.1071/rdv31n1ab128 ◽

2019 ◽

Vol 31 (1) ◽

pp. 189

Author(s):

F. Navarrete ◽

E. Mellisho ◽

Y. Wang ◽

J. Cabezas ◽

L. L. Rodriguez-Alvarez ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Gene Ontology ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Cell Types ◽

Biological Traits ◽

Next Generation ◽

Pca Analysis ◽

Heat Map

Earlier we successfully isolated and characterised endometrial (eMSC) and adipose (aMSC) mesenchymal stem cells from the same donors. Mesenchymal stem cells share biological traits but display different surface marker phenotype and migration ability. Here we extended our research to their mRNA signature using next-generation sequencing. The RNA from cells (3 biological replicates from each cell type and 3 technical replicates) at 90% confluence was extracted using a total RNA extraction kit and sent for mRNA-Seq (Norgen, Ontario, Canada; Illumina Sequencing Platform NextSEqn 500). Raw 76-bp single-end reads were aligned against the EquCab3 genome using RNA-STAR aligner. Counts were filtrated at a minimum of 5. Pairwise comparisons between the cell types were the input for gene ontology enrichment analysis. Only genes differentially expressed (DE) with 5 folds change (FC; P<0.05) were analysed. For DE analysis, eMSC were set as control and compared with aMSC. Unsupervised hierarchical clustering of the global gene expression signatures was done to compare the samples from each line using principal component analysis (PCA) and EdgeR: v3.20.9. Gene expression was normalized using FPKM. The heat map was built using R studio with G-plot package. A total of 14,896 transcripts with at least 5 reads were found; of these, 1598 were DE: 627 up-regulated (FC range: 2 to 236×) and 971 down-regulated (FC range: 2 to 464×) in eMSC. There was a marked dispersion in the FC of up- and down-regulated genes (>50×: 8 and 13; >20×, <50×: 9 and 17; >10×, <20×: 29 and 63; >5×, <10×: 91 and 130 and >2×, <5×: 490 and 748, respectively). Only genes DE with FC at least 5× were used for gene ontology and PCA analysis. Though 14,058 genes were common to both cell types, specific set of genes were found only in eMSC (n=162) or aMSC (n=676). Among the top 50 genes overexpressed in eMSC, several genes key for stem cell growth, immune response, migration and angiogenesis were found: TRIL, CXCL8, PDGF-D, SEMA5A, PTGS2, FGD, LAMA2, IL36G. In the top 50 down-regulated genes, some pivotal for osteoblast, adipogenic and neural differentiation were dramatically down-regulated (GPM6B, SCARA5 and NOTCH3 or NEFM, respectively), but no genes involved in immune rejection or stem cell proliferation were found. In gene ontology, the categories represented the most were cellular, developmental, metabolic, and immune system processes, as well as biological regulation, response to stimuli, organellar biogenesis, locomotion, localization and biological adhesion. Heat map and PCA analysis showed that one individual cell line from each type diverged markedly from the shared pattern. Individual variability of the donors may impinge upon the results; nevertheless, striking differences in the mRNA portfolio of eMSC and aMSC were detected. The importance and potential biological role of several of the genes and processes named above will be discussed in detail elsewhere. This work was supported by grant FONDECYT REGULAR 1150757 and the Government of Chile.

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Comparison of the effect of cigarette smoke on mesenchymal stem cells and dental stem cells

AJP Cell Physiology ◽

10.1152/ajpcell.00217.2020 ◽

2020 ◽

Author(s):

Brandon Nguyen ◽

Tamer Alpagot ◽

Heesoo Oh ◽

David Ojcius ◽

Nan Xiao

Keyword(s):

United States ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Regenerative Medicine ◽

Cigarette Smoking ◽

Cigarette Smoke ◽

The United States ◽

Systemic Effects ◽

Dental Stem Cells ◽

The Impact

The persistent prevalence of cigarette smoking continues to contribute to preventable disease and death in the United States. Although much is known about the deleterious systemic effects of cigarette smoke and nicotine, some clinically relevant areas still remain unclear, such as the impact of cigarette smoke and nicotine on stem cells and the subsequent implications in regenerative medicine. This review focuses on recent studies on the effect of cigarette smoke and one of its deleterious components nicotine on mesenchymal stem cells, with an emphasis on dental stem cells.

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The Effect of Blood-Derived Products on the Chondrogenic and Osteogenic Differentiation Potential of Adipose-Derived Mesenchymal Stem Cells Originated from Three Different Locations

Stem Cells International ◽

10.1155/2019/1358267 ◽

2019 ◽

Vol 2019 ◽

pp. 1-20 ◽

Cited By ~ 1

Author(s):

Markus Neubauer ◽

Olga Kuten ◽

Christoph Stotter ◽

Karina Kramer ◽

Andrea De Luna ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Regenerative Medicine ◽

Blood Product ◽

Blood Products ◽

Fat Tissue ◽

Differentiation Potential ◽

Lower Blood

Background. Adipose-derived mesenchymal stem cells (AD-MSCs) from fat tissue considered “surgical waste” during joint surgery may provide a potent source for regenerative medicine. Intra-articular, homologous fat tissue (Hoffa’s fat pad, pouch fat) might possess a superior chondrogenic and osteogenic differentiation potential in comparison to extra-articular, nonhomologous fat. Blood products might further enhance this potential. Methods. AD-MSCs were isolated from fat tissue of 3 donors from 3 locations each, during total knee replacement. Isolated cells were analyzed via flow cytometry. Cells were supplemented with blood products: two types of platelet-rich plasma (EPRP—PRP prepared in the presence of EDTA; CPRP—PRP prepared in the presence of citrate), hyperacute serum (hypACT), and standard fetal calf serum (FCS) as a positive control. The viability of the cells was determined by XTT assay, and the progress of differentiation was tested via histological staining and monitoring of specific gene expression. Results. Blood products enhance ex vivo cell metabolism. Chondrogenesis is enhanced by EDTA-PRP and osteogenesis by citrate PRP, whereas hyperacute serum enhances both differentiations comparably. This finding was consistent in histological analysis as well as in gene expression. Lower blood product concentrations and shorter differentiation periods lead to superior histological results for chondrogenesis. Both PRP types had a different biological effect depending upon concentration, whereas hyperacute serum seemed to have a more consistent effect, independent of the used concentration. Conclusion. (i) Blood product preparation method, (ii) type of anticoagulant, (iii) differentiation time, and (iv) blood product concentration have a significant influence on stem cell viability and the differentiation potential, favouring no use of anticoagulation, shorter differentiation time, and lower blood product concentrations. Cell-free blood products like hyperacute serum may be considered as an alternative supplementation in regenerative medicine, especially for stem cell therapies.

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Preparing Sheep Bladder Scaffold and Examining the Differentiation of Mesenchymal Stem Cell into Myocytes on Scaffolds

Gene Cell and Tissue ◽

10.5812/gct.114576 ◽

2021 ◽

Vol In Press (In Press) ◽

Author(s):

Reza Najafi ◽

Asadollah Asadi ◽

Saber Zahri ◽

Arash Abdolmaleki

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Maximum Level ◽

Specific Gene ◽

Biological Scaffolds ◽

Marrow Mesenchymal Stem Cells ◽

Physical And Chemical

Background: Tissue engineering may be used to repair, preserve, or improve tissues and organs. In this regard, acellular biological scaffolds are mainly used to reconstruct damaged tissues in regenerative medicine. Objectives: The present study examined the in vitro process of myocytes differentiated from bone marrow mesenchymal stem cells (BM‐MSCs) on the sheep bladder scaffold induced by 5-azacytidine. Methods: Decellularization was performed using a mixed method (physical and chemical) to prepare scaffolds kept at -20°C. The 5-azacytidine was used to induce BM‐MSCs to myocytes. Moreover, the muscle-specific gene expression (Desmin, α-Actinin, Myo D) was evaluated using the RT-PCR method. Results: It was revealed that BM‐MSCs on the scaffold had high proliferation and differentiation potentials. Desmin and α-Actinin gene expression marked the differentiation at the end of the fourth week. Moreover, the results of Masson’s trichrome staining at the end of the second, third and, fourth weeks also indicated that the first differentiation signs emerged at the end of the second week. Furthermore, differentiation reached its maximum level during the fourth week. Conclusions: According to the findings, combining physical and chemical methods was the best technique to prepare the bladder scaffold so that the bone marrow mesenchymal stem cells can be differentiated into myocytes on the bladder scaffold affected by 5-azacytidine (5 µmol), and As the induction time increases to day 28, myocyte cells become more developed.

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