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

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

Catalysts ◽

10.3390/catal11010062 ◽

2021 ◽

Vol 11 (1) ◽

pp. 62

Author(s):

Won-Yong Jeon ◽

Seyoung Mun ◽

Wei Beng Ng ◽

Keunsoo Kang ◽

Kyudong Han ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Electrical Stimulation ◽

Regenerative Medicine ◽

Cell Morphology ◽

Electrical Current ◽

Specific Gene ◽

Next Generation ◽

The Impact

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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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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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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PPIA and YWHAZ Constitute a Stable Pair of Reference Genes during Electrical Stimulation in Mesenchymal Stem Cells

Applied Sciences ◽

10.3390/app12010153 ◽

2021 ◽

Vol 12 (1) ◽

pp. 153

Author(s):

Lynsey Steel ◽

David M. Ansell ◽

Enrique Amaya ◽

Sarah H. Cartmell

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Electrical Stimulation ◽

Reference Gene ◽

Reference Genes ◽

Adult Stem Cells ◽

Expression Studies ◽

Proliferation And Differentiation ◽

Gene Expression Studies

Mesenchymal stem cells (MSCs) are multipotent adult stem cells with great potential in regenerative medicine. One method for stimulating proliferation and differentiation of MSCs is via electrical stimulation (ES). A valuable approach for evaluating the response of MSCs to ES is to assess changes in gene expression, relative to one or more reference genes. In a survey of 25 publications that used ES on cells, 70% selected GAPDH as the reference gene. We conducted a study to assess the suitability of six potential reference genes on an immortalized human MSC line following direct current ES at seeding densities of 5000 and 10,000 cells/cm2. We employed three methods to validate the most stable reference genes from qRT-PCR data. Our findings show that GAPDH and ACTB exhibit reduced stability when seeded at 5000 cell/cm2. In contrast, we found that the most stable genes across both plating densities and stimulation regimes were PPIA and YWHAZ. Thus, in ES gene expression studies in MSCs, we support the use of PPIA and YWHAZ as an optimal reference gene pair, and discourage the use of ACTB and GAPDH at lower seeding densities. However, it is strongly recommended that similar verification studies are carried out based on cell type and different ES conditions.

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A Review of the Impact of Electrical Stimulation on the Stem Cells Fate and Its Application in Regenerative Medicine and Cancer Treatment

Journal of Cell & Tissue ◽

10.52547/jct.11.2.139 ◽

2020 ◽

Vol 11 (2) ◽

pp. 139-153

Author(s):

AR Farmani ◽

M Mohammad Salehi ◽

F Mahdavinezhad ◽

M Kouhestani ◽

S Mohammadi ◽

...

Keyword(s):

Stem Cells ◽

Electrical Stimulation ◽

Regenerative Medicine ◽

Cancer Treatment ◽

The Impact

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From Human Mesenchymal Stem Cells to Insulin-Producing Cells: Comparison between Bone Marrow- and Adipose Tissue-Derived Cells

BioMed Research International ◽

10.1155/2017/3854232 ◽

2017 ◽

Vol 2017 ◽

pp. 1-9 ◽

Cited By ~ 13

Author(s):

Mahmoud M. Gabr ◽

Mahmoud M. Zakaria ◽

Ayman F. Refaie ◽

Engy A. Abdel-Rahman ◽

Asmaa M. Reda ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Regenerative Medicine ◽

C Peptide ◽

Metabolic Characteristics ◽

Insulin Producing Cells ◽

Glucose Challenge

The aim of this study is to compare human bone marrow-derived mesenchymal stem cells (BM-MSCs) and adipose tissue-derived mesenchymal stem cells (AT-MSCs), for their differentiation potentials to form insulin-producing cells. BM-MSCs were obtained during elective orthotopic surgery and AT-MSCs from fatty aspirates during elective cosmetics procedures. Following their expansion, cells were characterized by phenotyping, trilineage differentiation ability, and basal gene expression of pluripotency genes and for their metabolic characteristics. Cells were differentiated according to a Trichostatin-A based protocol. The differentiated cells were evaluated by immunocytochemistry staining for insulin and c-peptide. In addition the expression of relevant pancreatic endocrine genes was determined. The release of insulin and c-peptide in response to a glucose challenge was also quantitated. There were some differences in basal gene expression and metabolic characteristics. After differentiation the proportion of the resulting insulin-producing cells (IPCs), was comparable among both cell sources. Again, there were no differences neither in the levels of gene expression nor in the amounts of insulin and c-peptide release as a function of glucose challenge. The properties, availability, and abundance of AT-MSCs render them well-suited for applications in regenerative medicine.Conclusion. BM-MSCs and AT-MSCs are comparable regarding their differential potential to form IPCs. The availability and properties of AT-MSCs render them well-suited for applications in regenerative medicine.

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