Reaching out: junctions between cardiac telocytes and cardiac stem cells in culture.

Telocytes (TCs) were previously shown by our group to form a tandem with stem/progenitor cells in cardiac stem cell (CSC) niches, fulfilling various roles in cardiac renewal. Among these, the ability to 'nurse' CSCs in situ, both through direct physical contact (junctions) as well as at a distance, by paracrine signalling or through extracellular vesicles containing mRNA. We employed electron microscopy to identify junctions (such as gap or adherens junctions) in a co-culture of cardiac TCs and CSCs. Gap junctions were observed between TCs, which formed networks, however, not between TCs and CSCs. Instead, we show that TCs and CSCs interact in culture forming heterocellular adherens junctions, as well as non-classical junctions such as puncta adherentia and stromal synapses. The stromal synapse formed between TCs and CSCs (both stromal cells) was frequently associated with the presence of electron-dense nanostructures (on average about 15 nm in length) connecting the two opposing membranes. The average width of the synaptic cleft was 30 nm, whereas the average length of the intercellular contact was 5 µm. Recent studies have shown that stem cells fail to adequately engraft and survive in the hostile environment of the injured myocardium, possibly as a result of the absence of the pro-regenerative components of the secretome (paracrine factors) and/or of neighbouring support cells. Herein, we emphasize the similarities between the junctions described in co-culture and the junctions identified between TCs and CSCs in situ. Reproducing a CSC niche in culture may represent a viable alternative to mono-cellular therapies.

Extracellular vesicles release by cardiac telocytes: electron microscopy and electron tomography.

Telocytes have been reported to play an important role in long-distance heterocellular communication in normal and diseased heart, both through direct contact (atypical junctions), as well as by releasing extracellular vesicles (EVs) which may act as paracrine mediators. Exosomes and ectosomes are the two main types of EVs, as classified by size and the mechanism of biogenesis. Using electron microscopy (EM) and electron tomography (ET) we have found that telocytes in culture release at least three types of EVs: exosomes (released from endosomes; 45 ± 8 nm), ectosomes (which bud directly from the plasma membrane; 128 ± 28 nm) and multivesicular cargos (MVC; 1 ± 0.4 µm), the latter containing tightly packaged endomembrane-bound vesicles (145 ± 35 nm). Electron tomography revealed that endomembrane vesicles are released into the extracellular space as a cargo enclosed by plasma membranes (estimated area of up to 3 µm(2)). This new type of EV, also released by telocytes in tissue, likely represents an essential component in the paracrine secretion of telocytes and may consequently be directly involved in heart physiology and regeneration.