Human adipose‐derived stromal/stem cells are distinct from dermal fibroblasts as evaluated by biological characterization and RNA sequencing

: The isolation and culture of murine bone marrow-derived mesenchymal stromal stem cells (mBMSCs) have attracted great interest in terms of the pre-clinical applications of stem cells in tissue engineering and regenerative medicine. In addition, culturing mBMSCs is important for studying the molecular mechanisms of bone remodelling using relevant transgenic mice. Several factors have created challenges in the isolation and high-yield expansion of homogenous mBMSCs; these factors include low frequencies of bone marrow-derived mesenchymal stromal stem cells (BMSCs) in bone marrow, variation among inbred mouse strains, contamination with haematopoietic progenitor cells (HPCs), the replicative senescence phenotype and cellular heterogeneity. In this review, we provide an overview of nearly all protocols used for isolating and culturing mBMSCs with the aim of clarifying the most important guidelines for culturing highly purified mBMSC populations retaining in vitro and in vivo differentiation potential.

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Hypoxic Culture Conditions for Mesenchymal Stromal/Stem Cells from Wharton’s Jelly: A Critical Parameter to Consider in a Therapeutic Context

Current Stem Cell Research & Therapy ◽

10.2174/1574888x09666140213204850 ◽

2014 ◽

Vol 9 (4) ◽

pp. 306-318 ◽

Cited By ~ 20

Author(s):

Loic Reppel ◽

Talar Margossian ◽

Layale Yaghi ◽

Philippe Moreau ◽

Nathalie Mercier ◽

...

Keyword(s):

Stem Cells ◽

Critical Parameter ◽

Culture Conditions ◽

Wharton’S Jelly ◽

Wharton's Jelly ◽

Therapeutic Context ◽

Stromal Stem Cells

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Single-cell RNA sequencing reveals adverse effects of paraquat on the fate commitment of murine neural stem cells

The Science of The Total Environment ◽

10.1016/j.scitotenv.2021.147386 ◽

2021 ◽

pp. 147386

Author(s):

Guiya Xiong ◽

Bing Zhang ◽

Bo Song ◽

Huan Luo ◽

Lina Zhao ◽

...

Keyword(s):

Stem Cells ◽

Adverse Effects ◽

Neural Stem Cells ◽

Single Cell ◽

Rna Sequencing ◽

Single Cell Rna Sequencing

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Skin regeneration is accelerated by a lower dose of multipotent mesenchymal stromal/stem cells—a paradigm change

Stem Cell Research & Therapy ◽

10.1186/s13287-020-02131-6 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Gertraud Eylert ◽

Reinhard Dolp ◽

Alexandra Parousis ◽

Richard Cheng ◽

Christopher Auger ◽

...

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Low Dose ◽

Dose Range ◽

High Dose ◽

Full Thickness ◽

Burn Wound ◽

Dermal Regeneration Template ◽

Yorkshire Pigs ◽

Stromal Stem Cells

Abstract Background Multipotent mesenchymal stromal/stem cell (MSC) therapy is under investigation in promising (pre-)clinical trials for wound healing, which is crucial for survival; however, the optimal cell dosage remains unknown. The aim was to investigate the efficacy of different low-to-high MSC dosages incorporated in a biodegradable collagen-based dermal regeneration template (DRT) Integra®. Methods We conducted a porcine study (N = 8 Yorkshire pigs) and seeded between 200 and 2,000,000 cells/cm2 of umbilical cord mesenchymal stromal/stem cells on the DRT and grafted it onto full-thickness burn excised wounds. On day 28, comparisons were made between the different low-to-high cell dose groups, the acellular control, a burn wound, and healthy skin. Result We found that the low dose range between 200 and 40,000 cells/cm2 regenerates the full-thickness burn excised wounds most efficaciously, followed by the middle dose range of 200,000–400,000 cells/cm2 and a high dose of 2,000,000 cells/cm2. The low dose of 40,000 cells/cm2 accelerated reepithelialization, reduced scarring, regenerated epidermal thickness superiorly, enhanced neovascularization, reduced fibrosis, and reduced type 1 and type 2 macrophages compared to other cell dosages and the acellular control. Conclusion This regenerative cell therapy study using MSCs shows efficacy toward a low dose, which changes the paradigm that more cells lead to better wound healing outcome.

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3D Functional Corneal Stromal Tissue Equivalent Based on Corneal Stromal Stem Cells and Multi-Layered Silk Film Architecture

PLoS ONE ◽

10.1371/journal.pone.0169504 ◽

2017 ◽

Vol 12 (1) ◽

pp. e0169504 ◽

Cited By ~ 32

Author(s):

Chiara E. Ghezzi ◽

Benedetto Marelli ◽

Fiorenzo G. Omenetto ◽

James L. Funderburgh ◽

David L. Kaplan

Keyword(s):

Stem Cells ◽

Tissue Equivalent ◽

Stromal Tissue ◽

Stromal Stem Cells ◽

Silk Film

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Identification of New Transcription Factors that Can Promote Pluripotent Reprogramming

Stem Cell Reviews and Reports ◽

10.1007/s12015-021-10220-z ◽

2021 ◽

Author(s):

Ping Huang ◽

Jieying Zhu ◽

Yu Liu ◽

Guihuan Liu ◽

Ran Zhang ◽

...

Keyword(s):

Stem Cells ◽

Transcription Factors ◽

Embryonic Stem Cells ◽

Rna Sequencing ◽

Human Urine ◽

Embryonic Stem ◽

Rna Seq ◽

Reprogramming Efficiency ◽

Yamanaka Factors ◽

Urine Cells

Abstract Background Four transcription factors, Oct4, Sox2, Klf4, and c-Myc (the Yamanka factors), can reprogram somatic cells to induced pluripotent stem cells (iPSCs). Many studies have provided a number of alternative combinations to the non-Yamanaka factors. However, it is clear that many additional transcription factors that can generate iPSCs remain to be discovered. Methods The chromatin accessibility and transcriptional level of human embryonic stem cells and human urine cells were compared by Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq) and RNA sequencing (RNA-seq) to identify potential reprogramming factors. Selected transcription factors were employed to reprogram urine cells, and the reprogramming efficiency was measured. Urine-derived iPSCs were detected for pluripotency by Immunofluorescence, quantitative polymerase chain reaction, RNA sequencing and teratoma formation test. Finally, we assessed the differentiation potential of the new iPSCs to cardiomyocytes in vitro. Results ATAC-seq and RNA-seq datasets predicted TEAD2, TEAD4 and ZIC3 as potential factors involved in urine cell reprogramming. Transfection of TEAD2, TEAD4 and ZIC3 (in the presence of Yamanaka factors) significantly improved the reprogramming efficiency of urine cells. We confirmed that the newly generated iPSCs possessed pluripotency characteristics similar to normal H1 embryonic stem cells. We also confirmed that the new iPSCs could differentiate to functional cardiomyocytes. Conclusions In conclusion, TEAD2, TEAD4 and ZIC3 can increase the efficiency of reprogramming human urine cells into iPSCs, and provides a new stem cell sources for the clinical application and modeling of cardiovascular disease. Graphical abstract

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Microfluidic Tools for Enhanced Characterization of Therapeutic Stem Cells and Prediction of Their Potential Antimicrobial Secretome

Antibiotics ◽

10.3390/antibiotics10070750 ◽

2021 ◽

Vol 10 (7) ◽

pp. 750

Author(s):

Pasquale Marrazzo ◽

Valeria Pizzuti ◽

Silvia Zia ◽

Azzurra Sargenti ◽

Daniele Gazzola ◽

...

Keyword(s):

Stem Cells ◽

Defense Mechanisms ◽

Live Cells ◽

Label Free ◽

Bacterial Diseases ◽

Prospective Application ◽

Therapy Strategies ◽

Stromal Stem Cells ◽

Different Sources

Antibiotic resistance is creating enormous attention on the development of new antibiotic-free therapy strategies for bacterial diseases. Mesenchymal stromal stem cells (MSCs) are the most promising candidates in current clinical trials and included in several cell-therapy protocols. Together with the well-known immunomodulatory and regenerative potential of the MSC secretome, these cells have shown direct and indirect anti-bacterial effects. However, the low reproducibility and standardization of MSCs from different sources are the current limitations prior to the purification of cell-free secreted antimicrobial peptides and exosomes. In order to improve MSC characterization, novel label-free functional tests, evaluating the biophysical properties of the cells, will be advantageous for their cell profiling, population sorting, and quality control. We discuss the potential of emerging microfluidic technologies providing new insights into density, shape, and size of live cells, starting from heterogeneous or 3D cultured samples. The prospective application of these technologies to studying MSC populations may contribute to developing new biopharmaceutical strategies with a view to naturally overcoming bacterial defense mechanisms.

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CD90low glioma-associated mesenchymal stromal/stem cells promote temozolomide resistance by activating FOXS1-mediated epithelial-mesenchymal transition in glioma cells

Stem Cell Research & Therapy ◽

10.1186/s13287-021-02458-8 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Bing-zhou Xue ◽

Wei Xiang ◽

Qing Zhang ◽

Hao-fei Wang ◽

Yu-jie Zhou ◽

...

Keyword(s):

Stem Cells ◽

Cell Line ◽

Epithelial Mesenchymal Transition ◽

Chemotherapy Resistance ◽

Glioma Cells ◽

Conditioned Media ◽

Cell Counting ◽

Mesenchymal Transition ◽

Mesenchymal Transformation ◽

Stromal Stem Cells

Abstract Background The tumour microenvironment contributes to chemotherapy resistance in gliomas, and glioma-associated mesenchymal stromal/stem cells (gaMSCs) are important stromal cell components that play multiple roles in tumour progression. However, whether gaMSCs affect chemotherapy resistance to the first-line agent temozolomide (TMZ) remains unclear. Herein, we explored the effect and mechanism of gaMSCs on resistance to TMZ in glioma cells. Methods Human glioma cells (cell line U87MG and primary glioblastoma cell line GBM-1) were cultured in conditioned media of gaMSCs and further treated with TMZ. The proliferation, apoptosis and migration of glioma cells were detected by Cell Counting Kit-8 (CCK-8), flow cytometry and wound-healing assays. The expression of FOXS1 in glioma cells was analysed by gene microarray, PCR and Western blotting. Then, FOXS1 expression in glioma cells was up- and downregulated by lentivirus transfection, and markers of the epithelial-mesenchymal transformation (EMT) process were detected. Tumour-bearing nude mice were established with different glioma cells and treated with TMZ to measure tumour size, survival time and Ki-67 expression. Finally, the expression of IL-6 in gaMSC subpopulations and its effects on FOXS1 expression in glioma cells were also investigated. Results Conditioned media of gaMSCs promoted the proliferation, migration and chemotherapy resistance of glioma cells. The increased expression of FOXS1 and activation of the EMT process in glioma cells under gaMSC-conditioned media were detected. The relationship of FOXS1, EMT and chemotherapy resistance in glioma cells was demonstrated through the regulation of FOXS1 expression in vitro and in vivo. Moreover, FOXS1 expression in glioma cells was increased by secretion of IL-6 mainly from the CD90low gaMSC subpopulation. Conclusions CD90low gaMSCs could increase FOXS1 expression in glioma cells by IL-6 secretion, thereby activating epithelial-mesenchymal transition and resistance to TMZ in glioma cells. These results indicate a new role of gaMSCs in chemotherapy resistance and provide novel therapeutic targets.

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