scholarly journals Optimized culture methods for isolating small extracellular vesicles derived from human induced pluripotent stem cells

2021 ◽  
Vol 10 (6) ◽  
Author(s):  
Ying Luo ◽  
Dunqin Gao ◽  
Peng Wang ◽  
Cheng Lou ◽  
Tong Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Extracellular Vesicles ◽  
Pluripotent Stem Cells ◽  
Culture Methods ◽  
Induced Pluripotent

Abstract 15466: Bioactive Contents and Cardiovascular Regenerative Potential of Extracellular Vesicles Derived From Induced Pluripotent Stem Cells

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Marta Adamiak ◽  
Sylwia Bobis-Wozowicz ◽  
Elzbieta Karnas ◽  
Sylwia Kedracka-Krok ◽  
Michal Sarna ◽  
...  
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Endothelial Cells ◽  
Induced Pluripotent Stem Cells ◽  
Real Time ◽  
Extracellular Vesicles ◽  
Pluripotent Stem Cells ◽  
Target Cells ◽  
Rt Pcr ◽  
Induced Pluripotent

Introduction: Extracellular vesicles (EVs) are tiny membrane-enclosed vesicles released by cells by membrane budding or exocytosis. The molecular profile of contents and regenerative potential of EVs derived from murine induced pluripotent stem cells (miPSCs) have not been investigated. Hypothesis: We hypothesized that miPSC-derived EVs carry bioactive substances, including mRNA and miRNA, which would influence the biology and fate of target cells. Methods: Murine iPSCs were generated using established protocols and EVs were isolated from culture supernatants by sequential centrifugation. Atomic force microscopy (AFM), flow cytometry, and real-time RT-PCR were used to characterize EV contents. Endothelial cells were exposed to miPSC-EVs in culture. Results: AFM and dynamic light scattering showed that miPSC-EVs are homogenous spherical vesicles smaller than 100 nm (Fig, A). High-sensitivity flow cytometry confirmed the presence of several iPSC- specific markers along with typical exosomal markers (CD9, CD63 and CD81) on miPSC-EVs. miPSC-EVs were enriched in mRNAs, miRNAs and proteins from donor iPSCs as shown by real-time RT-PCR and mass spectroscopy, respectively. Moreover, miPSC-EVs contained transcripts regulating pluripotency, self-renewal and differentiation, including Oct-4, Nanog, GATA-4 as well as miR290-295 cluster (Fig, B,C). Importantly, several miRNAs were found to be higher in EVs when compared with parental miPSCs (Fig, B,C). Endothelial cells treated with miPSC-EVs exhibited greater proliferative, metabolic and angiogenic activities, and were more resistant to apoptosis. Conclusions: Our data show, for the first time, that miPSC-derived EVs are natural nanocarriers capable of transferring bioactive contents to mature cells affecting target cell function as well as regenerative potential. We conclude that iPSC-EVs may represent safe therapeutic alternatives to whole cell-based therapy for cardiovascular repair.

scholarly journals miR-212/132-Enriched Extracellular Vesicles Promote Differentiation of Induced Pluripotent Stem Cells Into Pancreatic Beta Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Chunyu Bai ◽  
Qiwei Ren ◽  
Haifeng Liu ◽  
Xiangchen Li ◽  
Weijun Guan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Beta Cell ◽  
Induced Pluripotent Stem Cells ◽  
Beta Cells ◽  
Extracellular Vesicles ◽  
Pluripotent Stem Cells ◽  
Pancreatic Beta Cells ◽  
Pancreatic Beta Cell ◽  
Induced Pluripotent

Pancreatic beta cell transplantation is the ideal method for treatment of type 1 diabetes mellitus (T1DM), and the generation of beta cells from induced pluripotent stem cells (iPSCs) of patients is a promising strategy. In this study, we improved a previous strategy to produce beta cells using extracellular vesicles (EVs) derived from mature beta cells and differentiated beta cells from iPSCs (i-Beta cells), which secreted insulin under glucose stimulation in vitro and ameliorated hyperglycemia in vivo. Mechanistic analyses revealed that EV-carried microRNA (miR)-212/132 (EV-miR-212/132) directly bound to the 3′ UTR of FBW7 to prevent its translation and FBW7 combined with NGN3 to accelerate its proteasomal degradation. EV-miR-212/132 stabilized NGN3 expression to promote differentiation of endocrine cells from induced iPSCs. Moreover, NGN3 bound to PDX1 to enhance transcription of endogenous miR-212/132 and formed a positive regulatory circuit that maintained the functions of mature pancreatic beta cells.ConclusionThis study describes a novel approach for beta cell production and supports the use of iPSCs for cell replacement therapy of T1DM.

scholarly journals Extracellular Vesicles from Skeletal Muscle Cells Efficiently Promote Myogenesis in Induced Pluripotent Stem Cells

Cells ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 1527 ◽  
Author(s):  
Denisa Baci ◽  
Maila Chirivì ◽  
Valentina Pace ◽  
Fabio Maiullari ◽  
Marika Milan ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Extracellular Vesicles ◽  
Pluripotent Stem Cells ◽  
Myogenic Differentiation ◽  
Bioactive Molecules ◽  
Patient Specific ◽  
Induced Pluripotent

The recent advances, offered by cell therapy in the regenerative medicine field, offer a revolutionary potential for the development of innovative cures to restore compromised physiological functions or organs. Adult myogenic precursors, such as myoblasts or satellite cells, possess a marked regenerative capacity, but the exploitation of this potential still encounters significant challenges in clinical application, due to low rate of proliferation in vitro, as well as a reduced self-renewal capacity. In this scenario, induced pluripotent stem cells (iPSCs) can offer not only an inexhaustible source of cells for regenerative therapeutic approaches, but also a valuable alternative for in vitro modeling of patient-specific diseases. In this study we established a reliable protocol to induce the myogenic differentiation of iPSCs, generated from pericytes and fibroblasts, exploiting skeletal muscle-derived extracellular vesicles (EVs), in combination with chemically defined factors. This genetic integration-free approach generates functional skeletal myotubes maintaining the engraftment ability in vivo. Our results demonstrate evidence that EVs can act as biological “shuttles” to deliver specific bioactive molecules for a successful transgene-free differentiation offering new opportunities for disease modeling and regenerative approaches.

scholarly journals Extracellular Vesicles Derived from Induced Pluripotent Stem Cells Promote Renoprotection in Acute Kidney Injury Model

Cells ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 453 ◽  
Author(s):  
Federica Collino ◽  
Jarlene A. Lopes ◽  
Marta Tapparo ◽  
Giovane G. Tortelote ◽  
Taís H. Kasai-Brunswick ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Death ◽  
Induced Pluripotent Stem Cells ◽  
Extracellular Vesicles ◽  
Pluripotent Stem Cells ◽  
Kidney Diseases ◽  
Ischemia Reperfusion ◽  
Gene Array ◽  
Wide Range ◽  
Induced Pluripotent

Induced pluripotent stem cells (iPSC) have been the focus of several studies due to their wide range of application, including in cellular therapy. The use of iPSC in regenerative medicine is limited by their tumorigenic potential. Extracellular vesicles (EV) derived from stem cells have been shown to support renal recovery after injury. However, no investigation has explored the potential of iPSC-EV in the treatment of kidney diseases. To evaluate this potential, we submitted renal tubule cells to hypoxia-reoxygenation injury, and we analyzed cell death rate and changes in functional mitochondria mass. An in vivo model of ischemia-reperfusion injury was used to evaluate morphological and functional alterations. Gene array profile was applied to investigate the mechanism involved in iPSC-EV effects. In addition, EV derived from adipose mesenchymal cells (ASC-EV) were also used to compare the potential of iPSC-EV in support of tissue recovery. The results showed that iPSC-EV were capable of reducing cell death and inflammatory response with similar efficacy than ASC-EV. Moreover, iPSC-EV protected functional mitochondria and regulated several genes associated with oxidative stress. Taken together, these results show that iPSC can be an alternative source of EV in the treatment of different aspects of kidney disease.

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