scholarly journals Deep Learning for Label-free Nuclei Detection from Implicit Phase Information of Mesenchymal Stem Cells

2021 ◽  
Author(s):  
zhengyun zhang ◽  
KIM WHYE LEONG ◽  
Krystyn Van Vliet ◽  
George Barbastathis ◽  
Andrea Ravasio
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Deep Learning ◽  
Label Free ◽  
Phase Information ◽  
Nuclei Detection

scholarly journals Label-Free Rapid Separation and Enrichment of Bone Marrow-Derived Mesenchymal Stem Cells from a Heterogeneous Cell Mixture Using a Dielectrophoresis Device

Sensors ◽  
2018 ◽  
Vol 18 (9) ◽  
pp. 3007 ◽  
Author(s):  
Junya Yoshioka ◽  
Yu Ohsugi ◽  
Toru Yoshitomi ◽  
Tomoyuki Yasukawa ◽  
Naoki Sasaki ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Gradient Method ◽  
Label Free ◽  
Rapid Separation ◽  
Isolation System ◽  
Tissue Samples ◽  
Cell Based Therapy

Bone marrow-derived mesenchymal stem cells (BMSCs) are an important cell resource for stem cell-based therapy, which are generally isolated and enriched by the density-gradient method based on cell size and density after collection of tissue samples. Since this method has limitations with regards to purity and repeatability, development of alternative label-free methods for BMSC separation is desired. In the present study, rapid label-free separation and enrichment of BMSCs from a heterogeneous cell mixture with bone marrow-derived promyelocytes was successfully achieved using a dielectrophoresis (DEP) device comprising saw-shaped electrodes. Upon application of an electric field, HL-60 cells as models of promyelocytes aggregated and floated between the saw-shaped electrodes, while UE7T-13 cells as models of BMSCs were effectively captured on the tips of the saw-shaped electrodes. After washing out the HL-60 cells from the device selectively, the purity of the UE7T-13 cells was increased from 33% to 83.5% within 5 min. Although further experiments and optimization are required, these results show the potential of the DEP device as a label-free rapid cell isolation system yielding high purity for rare and precious cells such as BMSCs.

scholarly journals UBA2 Activates Wnt/ β-catenin Signaling Pathway during Protection of R28 Retinal Precursor Cells from Hypoxia by Extracellular Vesicles derived from Placental Mesenchymal Stem Cells

2020 ◽  
Author(s):  
Kyungmin Koh ◽  
Mira Park ◽  
Eun Soo Bae ◽  
Van-An Duong ◽  
Jong-Moon Park ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Treatment Group ◽  
Signaling Pathway ◽  
Precursor Cells ◽  
Therapeutic Effects ◽  
Label Free ◽  
Related Factors ◽  
Cell Therapies ◽  
Placental Mesenchymal Stem Cells

Abstract Background: Stem cell transplantation has been proposed as an alternative treatment for intractable optic nerve disorders characterized by irrecoverable loss of cells. Mesenchymal stem cells, with varying tissue regeneration and recovery capabilities, are being considered for potential cell therapies. To overcome the limitations of cell therapy, we isolated exosomes from human placenta–derived mesenchymal stem cells (hPMSCs), and investigated their therapeutic effects in R28 cells (retinal precursor cells) exposed to CoCl2. Method: After nine hours of exposure to CoCl2, the hypoxic damaged R28 cells were divided into non treatment group (CoCl2+R28 cells) and treatment group (CoCl2+R28 cells treated with exosome). Immunoblot analysis was performed for Pcna, Hif-1α, Vegf, Vimentin, Thy-1, Gap43, Ermn, Neuroflament, Wnt3a, β-catenin, phospo-GSK3β, Lef-1, UBA2, Skp1, βTrcp, and ubiquitin. The proteomes of each group were analyzed by liquid chromatography/tandem mass (LC-MS/MS) spectrometry. Differentially expressed proteins (DEPs) were detected by label-free quantification and the interactions of the proteins were examined through signal transduction pathway and gene ontology analysis. Result: We observed that Exosome could significantly recover proliferation damaged by CoCl2 treatment. In addition, treatment group presented the decreased expression of Hif-1α protein (P < 0.05) and increased expression of proliferation marker, Pcna, and nerve regeneration–related factors such as Vimentin, Thy-1 and Neuroflament (P < 0.05) compared with non-treatment group. In total, 200 DEPs were identified in non-treatment group and treatment group (fold change ≥ 2, p < 0.05). Catenin and ubiquitin systems (UBA2, UBE2E3, UBE2I) were found in both the DEP lists of downregulated proteins from non-treatment group and upregulated proteins from treatment group. The mRNA expressions of ubiquitin systems were significantly decreased under hypoxic condition. Moreover, UBA2 and Wnt/β-catenin protein were associated with rescue of the hypoxic damaged R28 cells. Using a siRNA system, we could find it out that hPMSC exosoms could not repair altered expressions of target proteins by CoCl2 in lacking UBA2 R28 cells. Conclusion: This study reported that hypoxic damaged expression of regeneration markers in R28 cells were significantly recovered by hPMSC exosomes. We could also demonstrate that UBA2 played a key role in activating the Wnt/β-catenin signaling pathway during protection of hypoxic damaged R28 cells, induced by hPMSC exosomes.

scholarly journals High-throughput and label-free isolation of senescent murine mesenchymal stem cells

2020 ◽  
Vol 14 (3) ◽  
pp. 034106
Author(s):  
Zhengkun Chen ◽  
Kuan Jiang ◽  
Zhou Zou ◽  
Xiaohe Luo ◽  
Chwee Teck Lim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
High Throughput ◽  
Label Free

Label-free mesenchymal stem cell enrichment from bone marrow samples by inertial microfluidics

2018 ◽  
Vol 10 (7) ◽  
pp. 713-721 ◽  
Author(s):  
Lap Man Lee ◽  
Jenna M. Rosano ◽  
Yi Wang ◽  
George J. Klarmann ◽  
Charles J. Garson ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Bone Marrow Aspirate ◽  
Label Free ◽  
Therapeutic Implications ◽  
Cartilage Reconstruction ◽  
Cell Enrichment ◽  
And Cartilage

Isolation of pure populations of mesenchymal stem cells from bone marrow aspirate is a critical need in regenerative medicine such as orthopedic and cartilage reconstruction with important clinical and therapeutic implications.

scholarly journals Comprehensive Proteomic Analysis of Exosomes Derived from Human Bone Marrow-, Adipose Tissue-, and Umbilical Cord- Mesenchymal Stem Cells

2020 ◽  
Author(s):  
Zhenggang Wang ◽  
Zhi-yi He ◽  
Shuang Liang ◽  
Qing Yang ◽  
Peng Cheng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Umbilical Cord ◽  
Regeneration Ability ◽  
Human Mscs ◽  
Label Free ◽  
Damage Repair ◽  
Potential Applications

Abstract Background: Mesenchymal stem cells (MSCs)-derived exosomes have shown comprehensive application prospects over the years. Despite performing similar functions, exosomes from different origins present heterogeneous characteristics and components; however, relative study remains scarce. Lacking of a valuable reference, researchers select source cells for exosome studies mainly based on accessibility and personal preference.Methods: In this study, exosomes secreted by MSCs derived from different tissues were isolated, by ultracentrifugation, and proteomics analysis was performed. A total of 1,014 proteins were detected using a label-free method. Results: Bioinformatics analysis revealed their shared function in the extracellular matrix receptor. Bone marrow-MSCs-derived exosomes showed superior regeneration ability, and adipose tissue-MSCs-derived exosomes played a significant role in immune regulation, whereas, umbilical cord-MSCs-derived exosomes were more prominent in tissue damage repair.Conclusions: This study systematically and comprehensively analyzes the human MSCs-derived exosomes via proteomics, which reveals their potential applications in different fields, so as to provide a reference for researchers to select optimal source cells in future exosome-related studies.

Long-term and label-free monitoring for osteogenic differentiation of mesenchymal stem cells using force sensor and impedance measurement

2020 ◽  
Vol 8 (43) ◽  
pp. 9913-9920
Author(s):  
Zhizhong Zhang ◽  
Tianyang Zheng ◽  
Rong Zhu
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Impedance Measurement ◽  
Traction Force ◽  
Force Sensor ◽  
Label Free ◽  
Cell Impedance ◽  
Cell Traction

Label-free and continuous multimodal measurements of cell traction force and cell impedance for studying osteogenic differentiation of stem cells.

scholarly journals UBA2 activates Wnt/β-catenin signaling pathway during protection of R28 retinal precursor cells from hypoxia by extracellular vesicles derived from placental mesenchymal stem cells

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Kyungmin Koh ◽  
Mira Park ◽  
Eun Soo Bae ◽  
Van-An Duong ◽  
Jong-Moon Park ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Treatment Group ◽  
Signaling Pathway ◽  
Precursor Cells ◽  
Therapeutic Effects ◽  
Label Free ◽  
Related Factors ◽  
Cell Therapies ◽  
Placental Mesenchymal Stem Cells

Abstract Background Stem cell transplantation has been proposed as an alternative treatment for intractable optic nerve disorders characterized by irrecoverable loss of cells. Mesenchymal stem cells, with varying tissue regeneration and recovery capabilities, are being considered for potential cell therapies. To overcome the limitations of cell therapy, we isolated exosomes from human placenta-derived mesenchymal stem cells (hPMSCs) and investigated their therapeutic effects in R28 cells (retinal precursor cells) exposed to CoCl2. Method After 9 h of exposure to CoCl2, the hypoxic damaged R28 cells were divided into the non-treatment group (CoCl2 + R28 cells) and treatment group (CoCl2 + R28 cells treated with exosome). Immunoblot analysis was performed for Pcna, Hif-1α, Vegf, Vimentin, Thy-1, Gap43, Ermn, Neuroflament, Wnt3a, β-catenin, phospo-GSK3β, Lef-1, UBA2, Skp1, βTrcp, and ubiquitin. The proteomes of each group were analyzed by liquid chromatography/tandem mass (LC-MS/MS) spectrometry. Differentially expressed proteins (DEPs) were detected by label-free quantification, and the interactions of the proteins were examined through signal transduction pathway and gene ontology analysis. Result We observed that exosome could significantly recover proliferation damaged by CoCl2 treatment. In addition, the treatment group presented the decreased expression of Hif-1α protein (P < 0.05) and increased expression of proliferation marker, Pcna, and nerve regeneration-related factors such as Vimentin, Thy-1, and Neuroflament (P < 0.05) compared with the non-treatment group. In total, 200 DEPs were identified in the non-treatment group and treatment group (fold change ≥ 2, p < 0.05). Catenin and ubiquitin systems (UBA2, UBE2E3, UBE2I) were found in both the DEP lists of downregulated proteins from the non-treatment group and upregulated proteins from the treatment group. The mRNA expressions of ubiquitin systems were significantly decreased under hypoxic conditions. Moreover, UBA2 and Wnt/β-catenin protein were associated with the rescue of the hypoxic damaged R28 cells. Using a siRNA system, we could find it out that hPMSC exosomes could not repair altered expressions of target proteins by CoCl2 in lacking UBA2 R28 cells. Conclusion This study reported that hypoxic damaged expression of regeneration markers in R28 cells was significantly recovered by hPMSC exosomes. We could also demonstrate that UBA2 played a key role in activating the Wnt/β-catenin signaling pathway during protection of hypoxic damaged R28 cells, induced by hPMSC exosomes.

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