scholarly journals A Novel Tension Machine Promotes Bone Marrow Mesenchymal Stem Cell Osteoblastic and Fibroblastic Differentiation by Applying Cyclic Tension

2021 ◽  
Vol 2021 ◽  
pp. 1-15
Author(s):  
Yi Zhao ◽  
Yiping Huang ◽  
Lingfei Jia ◽  
Ruoxi Wang ◽  
Kuang Tan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Cyclic Tension ◽  
Marrow Mesenchymal Stem Cells ◽  
Tension Time ◽  
Mrna And Protein Expression ◽  
Cell Processes ◽  
Polymerase Chain

Bone marrow mesenchymal stem cells (BMSCs) are intraosseous stem cells, and the effects of tensile strain on BMSC differentiation mediate several bone-related treatments. To study the response of BMSCs under tension, we designed and developed a small cellular tension instrument, iStrain. When iStrain applied tension on BMSCs, these cells exhibited convergence in the alignment direction and lengthening of the cell processes and cell body. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting demonstrated that iStrain-mediated cyclic tension promotes the differentiation of BMSCs toward osteogenesis and fibrogenesis. And the mRNA and protein expression of differentiation-related genes changes with the extension of tension time.

scholarly journals SiglecF(HI) Marks Late‐Stage Neutrophils of the Infarcted Heart: A Single‐Cell Transcriptomic Analysis of Neutrophil Diversification

2021 ◽  
Vol 10 (4) ◽  
Author(s):  
David M. Calcagno ◽  
Claire Zhang ◽  
Avinash Toomu ◽  
Kenneth Huang ◽  
Van K. Ninh ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Single Cell ◽  
Negative Selection ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Cell Sorter ◽  
Infarcted Heart ◽  
Single Cell Rna Sequencing ◽  
Polymerase Chain ◽  
Deficient Mice

Background Neutrophils are thought to be short‐lived first responders to tissue injuries such as myocardial infarction (MI), but little is known about their diversification or dynamics. Methods and Results We permanently ligated the left anterior descending coronary arteries of mice and performed single‐cell RNA sequencing and analysis of >28 000 neutrophil transcriptomes isolated from the heart, peripheral blood, and bone marrow of mice on days 1 to 4 after MI or at steady‐state. Unsupervised clustering of cardiac neutrophils revealed 5 major subsets, 3 of which originated in the bone marrow, including a late‐emerging granulocyte expressing SiglecF, a marker classically used to define eosinophils. SiglecF HI neutrophils represented ≈25% of neutrophils on day 1 and grew to account for >50% of neutrophils by day 4 post‐MI. Validation studies using quantitative polymerase chain reaction of fluorescent‐activated cell sorter sorted Ly6G + SiglecF HI and Ly6G + SiglecF LO neutrophils confirmed the distinct nature of these populations. To confirm that the cells were neutrophils rather than eosinophils, we infarcted GATA‐deficient mice (∆dblGATA) and observed similar quantities of infiltrating Ly6G + SiglecF HI cells despite marked reductions of conventional eosinophils. In contrast to other neutrophil subsets, Ly6G + SiglecF HI neutrophils expressed high levels of Myc‐regulated genes, which are associated with longevity and are consistent with the persistence of this population on day 4 after MI. Conclusions Overall, our data provide a spatial and temporal atlas of neutrophil specialization in response to MI and reveal a dynamic proinflammatory cardiac Ly6G + SigF + (Myc + NFϰB + ) neutrophil that has been overlooked because of negative selection.

scholarly journals MicroRNA-425-5p modulates osteoporosis by targeting annexin A2

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Guanghua Chen ◽  
Guizhi Huang ◽  
Han Lin ◽  
Xinyou Wu ◽  
Xiaoyan Tan ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Bone Marrow ◽  
Cell Apoptosis ◽  
Chain Reaction ◽  
Potential Therapeutic Target ◽  
Treatment Of Osteoporosis ◽  
Qrt Pcr ◽  
Proliferation And Differentiation ◽  
Marrow Mesenchymal Stem Cells ◽  
Polymerase Chain

Abstract Background Studies have shown that the decrease of osteogenic differentiation of bone marrow mesenchymal stem cells (MSC) is an important mechanism of osteoporosis. The object of this study was to explore the role and mechanism of microRNA miR-425-5p in the differentiation of MSC. Methods The expression of miR-425-5p in MSC was detected by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Cell proliferation, cell cycle and apoptosis were detected by CCK-8 colorimetry and flow cytometry. The expression of TNF were detected by ELISA. Results Our data show that MiR-425-5p could modulate TNF-induced cell apoptosis, proliferation, and differentiation. ANXA2 is also the target of miR-425-5p and ANXA2 was involved in TNF-induced MSC cell apoptosis, proliferation, and differentiation. In addition, MiR-425-5p enhanced osteoporosis in mice. Conclusion MiR-425-5p might serve as a potential therapeutic target for the treatment of osteoporosis.

scholarly journals Lentiviral-mediated ephrin B2 gene modification of rat bone marrow mesenchymal stem cells

2019 ◽  
Vol 47 (7) ◽  
pp. 3282-3298
Author(s):  
Min Zhu ◽  
Yu Hua ◽  
Jian Tang ◽  
Xiaoke Zhao ◽  
Ling Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Neuronal Cell ◽  
Protein Levels ◽  
Marrow Mesenchymal Stem Cells ◽  
Lentivirus Vectors ◽  
Migratory Cells ◽  
Rat Bone

Objective To determine the effect of the upregulation or knockdown of the ephrinB2 ( Efnb2) gene and the effect of EphB4/EphrinB2 signalling in rat bone marrow mesenchymal stem cells (BMSCs). Methods Rat BMSCs were infected with lentivirus vectors carrying EphrinB2 and shRNA-EphrinB2. EphrinB2 mRNA and protein levels were quantified. At 28 days of culture with neuronal cell-conditioned differentiation medium, levels of microtubule-associated protein 2 (MAP2), CD133 and nestin were detected in EphrinB2/BMSCs and shEphrinB2/BMSCs using quantitative polymerase chain reaction and immunofluorescence. The ability of these cells to migrate was evaluated using a transwell assay. Results BMSCs were successfully isolated as indicated by their CD90+ CD29+ CD34– CD45– phenotype. Three days after ephrinB2 transduction, BMSC cell bodies began to shrink and differentiate into neuron-like cells. At 28 days, levels of MAP2, CD133 and nestin, as well as the number of migratory cells, were higher in lenti-EphrinB2-BMSCs than in the two control groups. The shEphrinB2/BMSCs had reduced levels of MAP2, CD133 and nestin; and a lower rate of cell migration. Similarly, increased levels of Grb4 andp21-activated kinase in the EphB4/EphrinB2 reverse signalling pathway were observed by Western blot. Conclusions LV-EphrinB2 can be efficiently transduced into BMSCs, which then differentiate into neuron-like cells.

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