scholarly journals Membrane potential (Vmem) measurements during mesenchymal stem cell (MSC) proliferation and osteogenic differentiation

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6341 ◽  
Author(s):  
Mit Balvantray Bhavsar ◽  
Gloria Cato ◽  
Alexander Hauschild ◽  
Liudmila Leppik ◽  
Karla Mychellyne Costa Oliveira ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Membrane Potential ◽  
Osteogenic Differentiation ◽  
Cell Function ◽  
Critical Role ◽  
Promote Cell Proliferation ◽  
Cell Behaviors ◽  
Cell Functions ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation

Background Electrochemical signals play an important role in cell communication and behavior. Electrically charged ions transported across cell membranes maintain an electrochemical imbalance that gives rise to bioelectric signaling, called membrane potential or Vmem. Vmem plays a key role in numerous inter- and intracellular functions that regulate cell behaviors like proliferation, differentiation and migration, all playing a critical role in embryonic development, healing, and regeneration. Methods With the goal of analyzing the changes in Vmem during cell proliferation and differentiation, here we used direct current electrical stimulation (EStim) to promote cell proliferation and differentiation and simultaneously tracked the corresponding changes in Vmem in adipose derived mesenchymal stem cells (AT-MSC). Results We found that EStim caused increased AT-MSC proliferation that corresponded to Vmem depolarization and increased osteogenic differentiation that corresponded to Vmem hyperpolarization. Taken together, this shows that Vmem changes associated with EStim induced cell proliferation and differentiation can be accurately tracked during these important cell functions. Using this tool to monitor Vmem changes associated with these important cell behaviors we hope to learn more about how these electrochemical cues regulate cell function with the ultimate goal of developing new EStim based treatments capable of controlling healing and regeneration.

Study on the Role of PML on Cell Proliferation and Differentiation In Human Mesenchymal Stem Cells

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4711-4711
Author(s):  
Jie Sun ◽  
He Huang ◽  
Weijun Zhong ◽  
Shan Fu
Keyword(s):  
Flow Cytometry ◽  
Cell Proliferation ◽  
Osteogenic Differentiation ◽  
Tumor Cells ◽  
Hela Cells ◽  
Human Mesenchymal Stem Cells ◽  
Rt Pcr ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Mrna And Protein Expression

Abstract Abstract 4711 Promyelocytic leukemia protein (PML) was found in discrete, punctuate subnuclear structures known as PML nuclear bodies (PML NBs) in normal somatic cells and is an important factor acting downstream of wnt/β-catenin in the regulation of cell proliferation and differentiation. It is well-known that PML can bind cbp/p300 to promote TCF/LEF dependent transcription of downstream genes in 293T cells. However, its function in human mesenchymal stem cells (hMSCs) remains unclear. In our former studies, we found PML NBs in mitosis metaphase of hMSCs were more abundant than in Hela cells and 293T cells, which indicates PML may have special functions in hMSCs compared with tumor cells and normal somatic cells. The aim of our study is to investigate PML's function during proliferation and differentiation in hMSCs in contrast to Hela cells and 293T cells. hMSCs were isolated from bone marrow of healthy volunteers by density gradient centrifugation using Ficoll-paque and cultured (LG-DMEM, 10%FBS) with their characteristic adherence and morphology. hMSC immunophenotype was analyzed by flow cytometry and demonstrated uniform positivity for CD29, CD44, CD166 and CD105, and negativity for CD34, CD45 and HLA-DR. MTT assay and flow cytometry were used to evaluate proliferation of hMSCs. Osteogenic differentiation of hMSCs was perfoprmed and differentiated cells were identified with biochemical and morphological approaches. The levels of PML mRNA and protein expression during proliferation and differentiation were detected by RT-PCR, western blot and immunofluorescence while Hela cells and 293T cells were used as controls. Results showed the mean levels of mRNA and protein expression significantly increased during proliferation of hMSCs and 293T cells and positively correlated with the proliferation status of hMSCs. However, the expression of PML decreased in proliferating Hela cells (Fig. 1A, B). Intranuclear NBs of MSCs immunostained with PML monoclonal antibody and showed increased levels during proliferation, but remained very low in Hela cells throughout their proliferation. (Fig2). The above results indicate PML may take part in regulating the proliferation of hMSCs, and the mechanism involved is different from that in tumor cells. In tumor cells, the main function of PML maybe a suppressor of malignant cell growth. And PML protein is reduced or almost completely lost by post-transcriptional mechanisms. This loss is associated with tumor cell proliferation while regulation of cell proliferation and differentiation is the more important function of PML in hMSCs. During osteogenic differentiation of MSCs, ALP which is an early marker of osteogenic differentiation increased while PML's expression accordingly increased as detected by RT-PCR. (Fig 1C). Intranuclear NBs during differentiation are bigger but unequal in size compared with controls. (Fig3) These changes indicated that PML may interact with other molecules in NBs and act as a complex to regulate the process of osteogneic differentiation, but the exact mechanism involved is unclear and needs further study. Disclosures: No relevant conflicts of interest to declare.

scholarly journals MicroRNA-19b-3p promotes cell proliferation and osteogenic differentiation of BMSCs by interacting with lncRNA H19

2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Gan Xiaoling ◽  
Liu Shuaibin ◽  
Liang Kailu
Keyword(s):  
Cell Proliferation ◽  
Protein Expression ◽  
Postmenopausal Osteoporosis ◽  
Critical Role ◽  
Dependent Manner ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
17Β Estradiol ◽  
Dose Dependent

Abstract Background To investigated the role of miR-19b-3p in regulating bone marrow mesenchymal stem cell (BMSC) proliferation and osteoblast differentiation. Methods The expression of miR-19b-3p and lncRNA H19 were measured in postmenopausal osteoporosis patients and BMP-22 induced BMSCs using qRT-PCR. MiR-19b-3p mimic or inhibitor was transfected into BMP-2 induced BMSCs. Cell proliferation was measured by BrdU method. Protein expression of RUNX2 and COL1A1 were measured by western blot. PcDNA3.1-lncRNA H19 with or without miR-19b-3p mimic was transfected into BMP-2 induced BMSCs. Results The expression of miR-19b-3p was significantly up-regulated in postmenopausal osteoporosis patients and BMP-2 induced BMSCs. MiR-19b-3p overexpression dramatically elevated, while miR-19b-3p inhibition decreased cell proliferation of BMSCs. Additionally, protein expression levels of RUNX2 and COL1A1, as well as ALP activity were significantly promoted by miR-19b-3p mimic transfection and inhibited by miR-19b-3p inhibitor transfection. LncRNA H19 was obviously down-regulated in postmenopausal osteoporosis patients. H19 overexpression significantly decreased cell proliferation and differentiation by down-regulating miR-19b-3p. Moreover, the expression of miR-19b-3p was inhibited, while H19 elvated in 17β-estradiol (E2) treated BMSCs in a dose-dependent manner. Conclusion These data were the first to reveal the critical role of H19/miR-19b-3p in postmenopausal osteoporosis, and provided a new therapeutic target for OP.

scholarly journals The Impact of Metallic Nanoparticles on Stem Cell Proliferation and Differentiation

Nanomaterials ◽  
2018 ◽  
Vol 8 (10) ◽  
pp. 761 ◽  
Author(s):  
Ahmed Abdal Dayem ◽  
Soo Lee ◽  
Ssang-Goo Cho
Keyword(s):  
Cell Proliferation ◽  
Stem Cell ◽  
Metallic Nanoparticles ◽  
Stem Cell Differentiation ◽  
Stem Cell Proliferation ◽  
Cell Functions ◽  
Cell Proliferation And Differentiation ◽  
Wide Range ◽  
Proliferation And Differentiation ◽  
The Impact

Nanotechnology has a wide range of medical and industrial applications. The impact of metallic nanoparticles (NPs) on the proliferation and differentiation of normal, cancer, and stem cells is well-studied. The preparation of NPs, along with their physicochemical properties, is related to their biological function. Interestingly, various mechanisms are implicated in metallic NP-induced cellular proliferation and differentiation, such as modulation of signaling pathways, generation of reactive oxygen species, and regulation of various transcription factors. In this review, we will shed light on the biomedical application of metallic NPs and the interaction between NPs and the cellular components. The in vitro and in vivo influence of metallic NPs on stem cell differentiation and proliferation, as well as the mechanisms behind potential toxicity, will be explored. A better understanding of the limitations related to the application of metallic NPs on stem cell proliferation and differentiation will afford clues for optimal design and preparation of metallic NPs for the modulation of stem cell functions and for clinical application in regenerative medicine.

scholarly journals The role of BMP6 in the proliferation and differentiation of chicken cartilage cells

2018 ◽  
Author(s):  
Fei Ye ◽  
Hengyong Xu ◽  
Huadong Yin ◽  
Xiaoling Zhao ◽  
Diyan Li ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Critical Role ◽  
Cartilage Cell ◽  
Differentiation Potential ◽  
Expression Levels ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Cartilage Cells ◽  
Collagen Ii

AbstractPrevious studies have indicated that bone morphogenetic protein (BMP) 6 plays an important role in skeletal system development and progression. However, the mechanism underlying the effects of BMP6 in cartilage cell proliferation and differentiation remains unknown. In this study, cartilage cells were isolated from shanks of chicken embryos and treated with different concentrations of GH. Cell proliferation and differentiation potential was assessed using real-time polymerase chain reaction (RT-PCR) and CCK-8 assays in vitro. The results showed that at 48 h, the Collagen II and BMP6 expression levels in 50 ng/μl GH-treated cartilage cells were significantly higher than in groups treated with 100 ng/μl or 200 ng/μl GH. We further observed that knockdown of BMP6 in cartilage cells led to significantly decreased expression levels of Collagen II and Collagen X. Moreover, the suppression of BMP6 expression by a specific siRNA vector led to significantly decreased expression levels of IGF1R, JAK, PKC, PTH, IHH and PTHrP. Taken together, our data suggest that BMP6 may play a critical role in chicken cartilage cell proliferation and differentiation through the regulation of IGF1, JAK2, PKC, PTH, and Ihh-PTHrP signaling pathways.

Icariin Stimulates hFOB 1.19 Osteoblast Proliferation and Differentiation via OPG/RANKL Mediated by the Estrogen Receptor

2020 ◽  
Vol 22 (1) ◽  
pp. 168-175 ◽  
Author(s):  
Lin-Jun Sun ◽  
Chong Li ◽  
Xiang-hao Wen ◽  
Lu Guo ◽  
Zi-Fen Guo ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Estrogen Receptor ◽  
Protein Expression ◽  
Chinese Herb ◽  
Human Osteoblast ◽  
Osteoblast Cells ◽  
Expression Ratio ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Mrna And Protein Expression

Background:: Icariin (ICA), one of the main effective components isolated from the traditional Chinese herb Epimedium brevicornu Maxim., has been reported to possess extensive pharmacological actions, including enhanced sexual function, immune regulation, anti-inflammation, and antiosteoporosis. Methods:: Our study was designed to investigate the effect of ICA on cell proliferation and differentiation and the molecular mechanism of OPG/RANKL mediated by the Estrogen Receptor (ER) in hFOB1.19 human osteoblast cells. Results:: The experimental results show that ICA can stimulate cell proliferation and increase the activity of Alkaline Phosphatase (ALP), Osteocalcin (BGP) and I Collagen (Col I) and a number of calcified nodules. Furthermore, the mRNA and protein expression of OPG and RANKL and the OPG/ RANKL mRNA and protein expression ratios were upregulated by ICA. The above-mentioned results indicated that the optimal concentration of ICA for stimulating osteogenesis was 50ng/mL. Subsequent mechanistic studies comparing 50ng/mL ICA with an estrogen receptor antagonist demonstrated that the effect of the upregulated expression is connected with the estrogen receptor. In conclusion, ICA can regulate bone formation by promoting cell proliferation and differentiation and upregulating the OPG/RANKL expression ratio by the ER in hFOB1.19 human osteoblast cells.

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