Abstract
A range of cell types, including embryonic stem cells, neurons and astrocytes have been shown to release extracellular vesicles (EVs) containing molecular cargo. Across cell types, EVs facilitate transfer of mRNA, microRNA and proteins between cells. Here we describe the release kinetics and content of EVs from mouse retinal progenitor cells (mRPCs). Interestingly, mRPC derived EVs contain mRNA, miRNA and proteins associated with multipotency and retinal development. Transcripts enclosed in mRPC EVs, include the transcription factors Pax6, Hes1, and Sox2, a mitotic chromosome stabilizer Ki67, and the neural intermediate filaments Nestin and GFAP. Proteomic analysis of EV content revealed retinogenic growth factors and morphogen proteins. mRPC EVs were shown to transfer GFP mRNA between cell populations. Finally, analysis of EV mediated functional cargo delivery, using the Cre-loxP recombination system, revealed transfer and uptake of Cre+ EVs, which were then internalized by target mRPCs activating responder loxP GFP expression. In summary, the data supports a paradigm of EV genetic material encapsulation and transfer within RPC populations. RPC EV transfer may influence recipient RPC transcriptional and post-transcriptional regulation, representing a novel mechanism of differentiation and fate determination during retinal development.
Highlights
A range of cell types, including embryonic stem cells, neurons and astrocytes have been shown to release extracellular vesicles (EVs) containing molecular cargo
Our results suggest that mouse retinal progenitor cells (mRPCs) EVs encapsulate miRNA, mRNA and proteins involved in multipotency and differentiation
A sample of Brownian motion exhibited by mRPC EVs in solution at 24 h is provided in Supplementary Video 1
Summary
A range of cell types, including embryonic stem cells, neurons and astrocytes have been shown to release extracellular vesicles (EVs) containing molecular cargo. MRPC EVs were shown to transfer GFP mRNA between cell populations. EVs include exosomes and microvesicles, which are lipid enclosed cell fragments with diameters ranging from approximately 30 nm to 1 μm, released from most cell. EVs enclose cytoplasmic and lipid bilayer embedded molecules, leading to encapsulation of unique combinations of microRNA, mRNA and proteins similar to those present in the cells from which they originate[17]. By transfer of mRNAs and proteins, EVs released from adult progenitor cells in kidney, lung and liver, induce de-differentiation of differentiated resident cells into stem cell-like phenotypes, leading to activation of regenerative programs[1,23]
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