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Sci Rep. 2016 Jan 22;6:19743. doi: 10.1038/srep19743.

Characterization of Induced Pluripotent Stem Cell Microvesicle Genesis, Morphology and Pluripotent Content.

Author information

1
Department of Biological Sciences, City University of New York, Lehman College, 250 Bedford Park Boulevard West, Bronx, NY 10468.
2
Biology Program, The Graduate School and University Center, City University of New York, 365 5th Avenue, New York, NY 10016.
3
Departments of Pediatrics, Hematology/Oncology Division, Weill Medical College of Cornell University, 413 E. 69th St., New York, NY 10021.
4
Biochemistry Doctoral Program, The Graduate School, City University of New York, New York, NY, 10468.
5
Department of Biological Sciences, City University of New York, Staten Island, 2800 Victory Boulevard, Staten Island, NY 10314.
6
Microenvironment and Metastasis Laboratory, Department of Molecular Oncology, Spanish National Cancer Research Centre (CNIO), Madrid, E28029, Spain.

Abstract

Microvesicles (MVs) are lipid bilayer-covered cell fragments that range in diameter from 30 nm-1 uM and are released from all cell types. An increasing number of studies reveal that MVs contain microRNA, mRNA and protein that can be detected in the extracellular space. In this study, we characterized induced pluripotent stem cell (iPSC) MV genesis, content and fusion to retinal progenitor cells (RPCs) in vitro. Nanoparticle tracking revealed that iPSCs released approximately 2200 MVs cell/hour in the first 12 hrs with an average diameter of 122 nm. Electron and light microscopic analysis of iPSCs showed MV release via lipid bilayer budding. The mRNA content of iPSC MVs was characterized and revealed the presence of the transcription factors Oct-3/4, Nanog, Klf4, and C-Myc. The protein content of iPSCs MVs, detected by immunogold electron microscopy, revealed the presence of the Oct-3/4 and Nanog. Isolated iPSC MVs were shown to fuse with RPCs in vitro at multiple points along the plasma membrane. These findings demonstrate that the mRNA and protein cargo in iPSC MVs have established roles in maintenance of pluripotency. Building on this work, iPSC derived MVs may be shown to be involved in maintaining cellular pluripotency and may have application in regenerative strategies for neural tissue.

PMID:
26797168
PMCID:
PMC4726265
DOI:
10.1038/srep19743
[Indexed for MEDLINE]
Free PMC Article

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