Telocytes in the Tumor Microenvironment.

There are several theories explaining the communication between cells in the context of tumor development. Over the years, interactions between normal and transformed cells have been observed. Generally, all types of cells make equal contributions to the formation of the tumor microenvironment - a location of primary oncogenesis. To date, several studies have reported the role of telocytes in cancer development, and many publications have emphasized the direct and indirect involvement of telocytes in angiogenesis; signaling through the secretion of extracellular vesicles, growth factors, and bioactive molecules; fibrosis development and extracellular matrix production; tissue repair and regeneration; and immune responses. Considering the main components of the tumor microenvironment, we will discuss the features of telocytes and their possible involvement in local tissue homeostasis.

Identification of Telocytes in the Pancreas of Turtles-A role in Cellular Communication.

The existence of telocytes (TCs) has not yet been established in the pancreases of aquatic reptiles. Here, we report TCs in the exocrine pancreas of Pelodiscus sinensis using transmission electron microscope (TEM), immunohistochemistry (IHC), and immunofluorescence (IF) techniques. TCs surrounded the acini and ducts of the connective tissue of the exocrine pancreas and between lobules and gland cells. The cells were located preferably close to the blood vessels, interlobular ducts, and nerve fibers. Ultrastructurally, TCs exhibited small and large bodies with thick and thin portions, podoms, and podomers, and prolongations that form dichotomous branching with hetero-cellular and homo-cellular junctions. The podom (thick) portions showed caveolae, mitochondria, rough endoplasmic reticulum, and vesicles. The nucleus carries heterochromatin and is irregular in shape. The shape of TCs depends on the number of telopodes (Tps) bearing long, short, spindle, triangular, and "beads on a string" shapes with twisted, tortuous prolongations and ramifications. Shed extracellular vesicles and exosomes were found frequently released from projections and Tps within connective tissue in the vicinity of the acini and collagen fibers. IHC and IF results showed CD34+, α-SMA+, and vimentin+, long and triangle-shaped TCs, consistent with the TEM findings. The presence of shaded vesicles from TCs might implicate their possible role in immune surveillance, tissue regeneration as well as regulatory functions in the reptilian pancreas.

The roles of telocytes in lung development and angiogenesis: An immunohistochemical, ultrastructural, scanning electron microscopy and morphometrical study.

Many studies have been carried out to investigate the occurrence and distribution of telocytes (TCs) in many organs. However, their morphological development is still unclear. This study was performed to demonstrate the morphological development of TCs in rabbits' lung from fetal to postnatal life using light-, electron- microscopy, immunohistochemistry, morphometrical and statistical analysis. During the fetal life, these cells formed an extensive network of telopodes (Tps) which were in close contact with developing alveoli, bronchioles, stem cells and many other interstitial components. In addition, the TCs' number was significantly increased around the neocapillaries in fetal lung. In the fetal life, TCs were stellate in shape and characterized by large cell bodies and many short Tps that contained abundant rER, mitochondria, and ribosomes. By gradual increasing of ages, TCs were spindle in shape with two Tps contained a massive amount of secretory structures (exosomes, ectosomes, and multivesicular bodies). Moreover, TCs in postnatal lung showed a significant decrease in number and diameter of their cell bodies and a significant increase in the length of Tps compared with those in fetal life. The TCs contributed with pneumocytes and endothelium in the formation of air-blood barrier. The TCs' immunohistochemical profiles for CD34, vimentin, c-kit, connexin 43, vascular endothelial growth factor (VEGF), and neuron- specific enolase (NSE) differed between ages during the lung development. This study provided an evidence that TCs contributed to angiogenesis, the formation of the air-blood barrier, lung organization, and development.

Telocytes in the architecture of uterine fibroids.

Knowledge of uterine fibroids has existed since the time of Hippocrates. However, there are still wide gaps in the understanding of its pathogenesis. No single theory explains the background of uterine fibroid pathology, which affects more than 50% of women worldwide. By contrast, a newly depicted cell type called telocytes was only recently identified in the past twenty years. These cells have evoked ambivalent opinions in the scientific community. The unique features of telocytes coupled with experimental evidence by numerous researchers and our hypotheses and conceptions are discussed in this review. We emphasize the main telocyte interactions in the context of the uterine fibroid architecture. This review reveals the pivotal role of telocytes, describing their contacts with smooth muscle cells, fibroblasts, vessels and nerves, inflammatory cells and stem cells. Our data are based on the latest publications and our own results.

Cardiac telocytes in normal and diseased hearts.

Our previous studies suggested that an important variable of the progression of contractile dysfunction to terminal heart failure is the imbalance between myocyte cell death and myocyte renewal. For this reason, preventing myocyte cell death and an increasing generation of new myocytes may represent attractive targets in the treatment of human heart failure. Prospective clues to enhance myocardial regeneration are the newly discovered cells termed telocytes, formerly called interstitial Cajal-like cells, which are believed to nurse or guide the endogenous and exogenous stem cells for activation and commitment, but they also act as supporting cells for progenitor cells migration toward injured myocardium. We have recently found that telocytes are reduced in the diseased and failing myocardium. Importantly, the imbalance between telocyte proliferation and telocyte death is responsible for the telocytes depletion in cardiac diseases leading to heart failure. We have also demonstrated that telocytes are influenced by the extracellular matrix protein composition such that the telocytes are almost absent in areas of severe fibrosis. It is plausible that the reduction in telocytes in diseased human hearts could participate in the abnormal three-dimensional spatial organization and disturbed intercellular signalling of the myocardium. Decreased telocytes in diseased hearts would also be predicted to alter the property of telocytes to maintain cardiac stem cell niche by decreasing the pool of cardiac stem cells and thereby impairing cardiac regeneration.

Telocytes in the reproductive organs: Current understanding and future challenges.

Over the past decades, we were witnessing spectacular molecular medicine advances. However, many of the reproductive medicine problems, such as fertility issues and premature birth still represent major challenges for obstetrics and gynecology worldwide. A new cell population - the telocytes (TCs) - were described in the interstitial space of many organs, and their possible implications in many important physiological and pathological processes should not be overlooked. In this article, we present a historical perspective outlining the landmarks in the discovery, evolution and achievements in the field of TCs over the last ten years. We focused on the potential roles of TCs in morphogenesis and maintenance of the normal three-dimensional architecture of tissues, in controlling of the stem cell microenvironment, as having anti-inflammatory and cancer-suppressing properties, participating in the immune surveillance, all mediated by direct homo- and heterocellular junctions or indirectly by extracellular vesicle release. Here, we overview the advances on TCs research in the reproductive organs (uterus and fallopian tube), accessory reproductive organs of female (mammary glands) and the temporary endocrine organ-placenta.

Telocytes: facts, speculations and myths (Review article).

Telocyte (TC) is an interstitial cell type with a small cellular body and extremely long tentacle-like extensions. TCs were discovered a decade ago and have specifc morphological characteristics, immunohistochemical and secretome profiles, electrophysiological properties, microRNA expression. Moreover, they are different in gene expression from other cells. TCs play an important role in plenty of processes. Apparently, they are involved in homeostasis, remodelling, regeneration, repair, embryogenesis, angiogenesis and even tumorigenesis. "Telocytes need the world", was emphasized by Professor Popescu and it will be actual at any time. This review summarizes particular features of TCs in different organs and systems, emphasizing their involvement in physiological and pathophysiological processes.

Telocytes in ileum of the Chinese giant salamander: ultrastructural evidence.

Telocytes (TCs) and their telopodes (Tps) have been found in various organs of many mammals, including in lower animals. However, knowledge of TCs in lower animals is still very limited. This study identified TCs and their Tps in the ileum of the Chinese giant salamander, Andrias davidianus (Amphibia: Caudata), by transmission electron microscopy. The TCs/Tps were found near epithelial cells, glandular cells and unmyelinated nerve fibres. Moreover, exosomes were also found to be present in between TCs/Tps and these cells.

Transmission electron microscope evidence of telocytes in canine dura mater.

Telocytes (TCs) are a novel type of interstitial cells present in a wide variety of organs and tissues (www.telocytes.com). Telocytes are identified morphologically by a small cell body and specific long prolongations (telopodes) alternating thin segments (podomers) with dilations (podoms). The presence of TCs in rat meninges has been identified in previous research. We here present further evidence that TCs existed in canine dura mater, closed to capillary and surrounded by a great deal of collagen fibres under transmission electron microscope.

Telocytes in female reproductive system (human and animal).

Telocytes (TCs) are a newly discovered type of cell with numerous functions. They have been found in a large variety of organs: heart (endo-, myo-, epi- and pericardium, myocardial sleeves, heart valves); digestive tract and annex glands (oesophagus, stomach, duodenum, jejunum, liver, gallbladder, salivary gland, exocrine pancreas); respiratory system (trachea and lungs); urinary system (kidney, renal pelvis, ureters, bladder, urethra); female reproductive system (uterus, Fallopian tube, placenta, mammary gland); vasculature (blood vessels, thoracic duct); serous membranes (mesentery and pleura); and other organs (skeletal muscle, meninges and choroid plexus, neuromuscular spindles, fascia lata, skin, eye, prostate, bone marrow). Likewise, TCs are widely distributed in vertebrates (fish, reptiles, birds, mammals, including human). This review summarizes particular features of TCs in the female reproductive system, emphasizing their involvement in physiological and pathophysiological processes.

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.

Telocytes of the human adult trigeminal ganglion.

Telocytes (TCs) are typically defined as cells with telopodes by their ultrastructural features. Their presence was reported in various organs, however little is known about their presence in human trigeminal ganglion. To address this issue, samples of trigeminal ganglia were tested by immunocytochemistry for CD34 and examined by transmission electron microscopy (TEM). We found that TCs are CD34 positive and form networks within the ganglion in close vicinity to microvessels and nerve fibers around the neuronal-glial units (NGUs). TEM examination confirmed the existence of spindle-shaped and bipolar TCs with one or two telopodes measuring between 15 to 53 µm. We propose that TCs are cells with stemness capacity which might contribute in regeneration and repair processes by: modulation of the stem cell activity or by acting as progenitors of other cells present in the normal tissue. In addition, further studies are needed to establish if they might influence the neuronal circuits.

Decoding Telocytes.

Telocytes (TCs) are a novel type of interstitial cells, with extremely long and thin cellular prolongations termed telopodes (Tps). TCs were first identified by Popescu et al. and described their finding as "cells with telopodes." The presence of TCs has been reported in the majority of tissues and organs (for details please visit www.telocytes.com ). TCs have been ignored or overlooked for a long time due to our inability to observe these cells via a light microscopy. TCs represent a distinct cell population, different from other types of interstitial cells, based on their distinct (ultra)structure, immunophenotype, microRNA profile, gene feature, proteome signature, and secretome features. As TCs have been suggested as new cellular targets for forthcoming therapies, developing specific methods to modulate TC numbers represents an important objective.

The Third Dimension of Telocytes Revealed by FIB-SEM Tomography.

Lately, spatial three-dimensional (3D) identity of cells and their interrelations with the environment that surrounds it represent a challenging trend with the purpose to achieve a holistic view over the functions. Combining data from different imaging of cells in the third dimension can offer insight into behavior modalities making a world of difference. This chapter outlines a breakthrough in telocyte research by volume electron microscopy with the aid of focused ion beam scanning electron microscopy (FIB-SEM). Reconstructing 3D (three-dimensional) appearance of telocytes from a set of two-dimensional (2D) images by FIB-SEM tomography allowed to extract valuable data about their volume in nanoscale dimensions such as the three-dimensional morphology of telopodes and extracellular vesicles.

Immunohistochemistry of Telocytes in the Uterus and Fallopian Tubes.

The seminal work of Popescu and colleagues first demonstrated the existence of a new cell type - the telocytes. We were among the first who reported the presence of such cells in the female genital tract and performed TEM examinations, as well as immunohistochemical staining in the attempt to find a specific marker. Telocytes from rat and from the human uterus and from human fallopian tube were extensively investigated initially by comparison with interstitial cells of Cajal. Progress in telocyte research led to the identification of different subtypes suggestive for a heterogeneous telocyte population which can even coexist in the same location. As a consequence, the functions of TCs are still elusive and can be considered a versatile phenomenon that depends on a variety of conditions, including signal reception and transmission of information via extracellular vesicles or by direct intercellular contact.

A Tale of Two Cells: Telocyte and Stem Cell Unique Relationship.

Telocytes have been identified as a distinctive type of interstitial cells and have been recognized in most tissues and organs. Telocytes are characterized by having extraordinary long cytoplasmic processes, telopodes, that extend to form three-dimensional networks and commonly constitute specialized forms of cell-to-cell junctions with other neighboring cells. Telocytes have been localized in the stem cell niche of different organs such as the heart, lung, skeletal muscle, and skin. Electron microscopy and electron tomography revealed a specialized link between telocytes and stem cells that postulates a potential role for telocytes during tissue regeneration and repair. In this review, the distribution of telocytes in different stem cell niches will be explored, highlighting the intimate relationship between the two types of cells and their possible functional relationship.

Telocytes and Their Extracellular Vesicles-Evidence and Hypotheses.

Entering the new millennium, nobody believed that there was the possibility of discovering a new cellular type. Nevertheless, telocytes (TCs) were described as a novel kind of interstitial cell. Ubiquitously distributed in the extracellular matrix of any tissue, TCs are regarded as cells with telopodes involved in intercellular communication by direct homo- and heterocellular junctions or by extracellular vesicle (EVs) release. Their discovery has aroused the interest of many research groups worldwide, and many researchers regard them as potentially regenerative cells. Given the experience of our laboratory, where these cells were first described, we review the evidence supporting the fact that TCs release EVs, and discuss alternative hypotheses about their future implications.

Skin telocytes versus fibroblasts: two distinct dermal cell populations.

It is already accepted that telocytes (TCs) represent a new type of interstitial cells in human dermis. In normal skin, TCs have particular spatial relations with different dermal structures such as blood vessels, hair follicles, arrector pili muscles or segments of sebaceous and/or eccrine sweat glands. The distribution and the density of TCs is affected in various skin pathological conditions. Previous studies mentioned the particular (ultra)structure of TCs and also their immunophenotype, miR imprint or proteome, genome or secretome features. As fibroblast is the most common intersitital cell (also in human dermis), a dedicated comparison between human skin TCs and fibroblasts (Fbs) was required to be performed. In this study, using different techniques, we document several points of difference between human dermis TCs and Fbs. By transmission electron microscopy (TEM) and scanning electron microscopy (SEM), we demonstrated TCs with their hallmark cellular prolongations - telopodes. Thus, we showed their ultrastructural distinctiveness from Fbs. By RayBio Human Cytokine Antibody Array V analyses performed on the supernatant from separately cultured TCs and Fbs, we detected the cytokine profile of both cell types, individually. Two of 79 detected cytokines - epithelial-derived neutrophil-activating peptide 78 and granulocyte chemotactic protein-2 - were 1.5 times higher in the supernatant of TCs (comparing with Fbs). On the other hand, 37 cytokines were at least 1.5 higher in Fbs supernatant (comparing with TCs), and among them six cytokines - interleukin 5, monocyte chemotactic protein-3 (MCP-3), MCP-4, macrophage inflammatory protein-3, angiogenin, thrombopoietin - being 9.5 times higher (results also confirmed by ELISA testing). In summary, using different techniques, we showed that human dermal TCs and Fbs are different in terms of ultrastructure and cytokine profile.

The failing human heart is characterized by decreased numbers of telocytes as result of apoptosis and altered extracellular matrix composition.

Telocytes (TCs) are a novel type of interstitial cells only recently described. This study aimed at characterizing and quantifying TCs and telopodes (Tps) in normal and diseased hearts. We have been suggested that TCs are influenced by the extracellular matrix (ECM) composition. We used transmission electron microscopy and c-kit immunolabelling to identify and quantify TCs in explanted human hearts with heart failure (HF) because of dilated, ischemic or inflammatory cardiomyopathy. LV myectomy samples from patients with aortic stenosis with preserved ejection fraction and samples from donor hearts which could not be used for transplantation served as controls. Quantitative immunoconfocal analysis revealed that 1 mm(2) of the normal myocardium contains 14.9 ± 3.4 TCs and 41.6 ± 5.9 Tps. As compared with the control group, the number of TCs and Tps in HF decreased more than twofold. There were no differences between HF and control in the number of Ki67-positive TCs. In contrast, terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling-positive TCs increased threefold in diseased hearts as compared to control. Significant inverse correlations were found between the amount of mature fibrillar collagen type I and the number of TCs (r = -0.84; P < 0.01) and Tps (r = -0.85; P < 0.01). The levels of degraded collagens showed a significant positive relationship with the TCs numbers. It is concluded that in HF the number of TCs are decreased because of higher rates of TCs apoptosis. Moreover, our results indicate that a close relationship exists between TCs and the ECM protein composition such that the number of TCs and Tps correlates negatively with the amount of mature fibrillar collagens and correlates positively with degraded collagens.

FIB-SEM tomography of human skin telocytes and their extracellular vesicles.

We have shown in 2012 the existence of telocytes (TCs) in human dermis. TCs were described by transmission electron microscopy (TEM) as interstitial cells located in non-epithelial spaces (stroma) of many organs (see www.telocytes.com). TCs have very long prolongations (tens to hundreds micrometers) named Telopodes (Tps). These Tps have a special conformation with dilated portions named podoms (containing mitochondria, endoplasmic reticulum and caveolae) and very thin segments (below resolving power of light microscopy), called podomers. To show the real 3D architecture of TC network, we used the most advanced available electron microscope technology: focused ion beam scanning electron microscopy (FIB-SEM) tomography. Generally, 3D reconstruction of dermal TCs by FIB-SEM tomography revealed the existence of Tps with various conformations: (i) long, flattened irregular veils (ribbon-like segments) with knobs, corresponding to podoms, and (ii) tubular structures (podomers) with uneven calibre because of irregular dilations (knobs) - the podoms. FIB-SEM tomography also showed numerous extracellular vesicles (diameter 438.6 ± 149.1 nm, n = 30) released by a human dermal TC. Our data might be useful for understanding the role(s) of TCs in intercellular signalling and communication, as well as for comprehension of pathologies like scleroderma, multiple sclerosis, psoriasis, etc.