Telocytes in regenerative medicine

Telocytes (TCs) are a distinct type of interstitial cells characterized by a small cell body and extremely long and thin telopodes (Tps). The presence of TCs has been documented in many tissues and organs (go to http://www.telocytes.com ). Functionally, TCs form a three‐dimensional (3D) interstitial network by homocellular and heterocellular communication and are involved in the maintenance of tissue homeostasis. As important interstitial cells to guide or nurse putative stem and progenitor cells in stem cell niches in a spectrum of tissues and organs, TCs contribute to tissue repair and regeneration. This review focuses on the latest progresses regarding TCs in the repair and regeneration of different tissues and organs, including heart, lung, skeletal muscle, skin, meninges and choroid plexus, eye, liver, uterus and urinary system. By targeting TCs alone or in tandem with stem cells, we might promote regeneration and prevent the evolution to irreversible tissue damage. Exploring pharmacological or non‐pharmacological methods to enhance the growth of TCs would be a novel therapeutic strategy besides exogenous transplantation for many diseased disorders.     

Proteome mapping of the Drosophila melanogaster male reproductive system

The fruit fly Drosophila melanogaster is an excellent model organism for studying insect reproductive biology. Although the gene expression profiles of both male and female reproductive organs have been studied in detail, their proteomic profiles and functional characteristics largely remained to be clarified. In this study, we conducted proteome mapping of the male internal reproductive organs using 2‐DE. We identified a total of 440 protein components from gels of the male reproductive organs (testis, seminal vesicle, accessory gland, ejaculatory duct, and ejaculatory bulb). A number of proteins associated with odorant/pheromone‐binding, lipid metabolism, proteolysis, and antioxidation were expressed tissue specifically in the male reproductive system. Based on our proteomic data set, we constructed reference proteome maps of the reproductive organs, which will provide valuable information toward a comprehensive understanding of Drosophila reproduction.     

A histological study of the accessory reproductive organs of female Loligo forbesi (Cephalopoda: Loliginidae)

In Loligo forbesi Steenstrup, the female reproductive system consists of the ovary and accessory reproductive organs which include the oviducal gland, the nidamental gland, the accessory nidamental gland and seminal receptacle. Histological studies were made on the accessory reproductive organs of female L. forbesi. The various changes observed during maturation are described and the functional significance discussed. The secretions produced by the oviducal gland and nidamental gland apparently form the egg coats. The seminal receptacle serves to store spermatozoa after mating. The function of the accessory nidamental gland is unknown.     

Cardiac telocytes are double positive for CD34/PDGFR‐α

Telocytes (TCs) are a distinct type of interstitial cells, which are featured with a small cellular body and long and thin elongations called telopodes (Tps). TCs have been widely identified in lots of tissues and organs including heart. Double staining for CD34/PDGFR‐β (Platelet‐derived growth factor receptor β) or CD34/Vimentin is considered to be critical for TC phenotyping. It has recently been proposed that CD34/PDGFR‐α (Platelet‐derived growth factor receptor α) is actually a specific marker for TCs including cardiac TCs although the direct evidence is still lacking. Here, we showed that cardiac TCs were double positive for CD34/PDGFR‐α in primary culture. CD34/PDGFR‐α positive cells (putative cardiac TCs) also existed in mice ventricle and human cardiac valves including mitral valve, tricuspid valve and aortic valve. Over 87% of cells in a TC‐enriched culture of rat cardiac interstitial cells were positive for PDGFR‐α, while CD34/PDGFR‐α double positive cells accounted for 30.25% of the whole cell population. We show that cardiac TCs are double positive for CD34/PDGFR‐α. Better understanding of the immunocytochemical phenotypes of cardiac TCs might help using cardiac TCs as a novel source in cardiac repair.     

The secretome of myocardial telocytes modulates the activity of cardiac stem cells

Telocytes (TCs) are interstitial cells that are present in numerous organs, including the heart interstitial space and cardiac stem cell niche. TCs are completely different from fibroblasts. TCs release extracellular vesicles that may interact with cardiac stem cells (CSCs) via paracrine effects. Data on the secretory profile of TCs and the bidirectional shuttle vesicular signalling mechanism between TCs and CSCs are scarce. We aimed to characterize and understand the in vitro effect of the TC secretome on CSC fate. Therefore, we studied the protein secretory profile using supernatants from mouse cultured cardiac TCs. We also performed a comparative secretome analysis using supernatants from rat cultured cardiac TCs, a pure CSC line and TCs‐CSCs in co‐culture using (i) high‐sensitivity on‐chip electrophoresis, (ii) surface‐enhanced laser desorption/ionization time‐of‐flight mass spectrometry and (iii) multiplex analysis by Luminex‐xMAP. We identified several highly expressed molecules in the mouse cardiac TC secretory profile: interleukin (IL)‐6, VEGF, macrophage inflammatory protein 1α (MIP‐1α), MIP‐2 and MCP‐1, which are also present in the proteome of rat cardiac TCs. In addition, rat cardiac TCs secrete a slightly greater number of cytokines, IL‐2, IL‐10, IL‐13 and some chemokines like, GRO‐KC. We found that VEGF, IL‐6 and some chemokines (all stimulated by IL‐6 signalling) are secreted by cardiac TCs and overexpressed in co‐cultures with CSCs. The expression levels of MIP‐2 and MIP‐1α increased twofold and fourfold, respectively, when TCs were co‐cultured with CSCs, while the expression of IL‐2 did not significantly differ between TCs and CSCs in mono culture and significantly decreased (twofold) in the co‐culture system. These data suggest that the TC secretome plays a modulatory role in stem cell proliferation and differentiation.     

Mating senescence and male reproductive organ size in the Mexican fruit fly

Ageing can reduce the probability that individuals reproduce. The present study investigates whether ageing influences the mating frequency of mass‐reared fertile and sterile Mexican fruit flies Anastrepha ludens (Loew). The ability of males of different ages to inhibit female remating is also determined, and the growth of male reproductive organs is measured as they age. Young males (6 days old) have a lower mating frequency than older males, and also have a lower capacity to inhibit female remating than older males. However, 7‐day‐old males are as likely to inhibit female remating as older males. Young males also have smaller reproductive organs than middle‐aged (21‐day‐old) or senescent males (57‐day‐old). These results have implications for the sterile insect technique because sterilized males of A. ludens are released in the field 6 days after emergence. The highest mating frequency, the lowest mating latency and the largest size of testes are observed at 21 days of age. Older males (57 days old) have more sperm in their seminal vesicles than young males (6 and 9 days old). Accessory glands take longer to grow to their complete size compared with testes, and mating frequency is more closely associated with accessory gland size than testes size. Furthermore, there are more sperm in the seminal vesicles during the afternoon period of peak sexual activity than during the morning when sexual activity is absent. These results indicate that, even at the onset of reproductive senescence, mass‐reared males of A. ludens are still capable of mating, as well as inhibiting remating in females.     

Ultrastructure damage of oviduct telocytes in rat model of acute salpingitis

Acute salpingitis (AS) is an inflammatory disease which causes severe damage to a subset of classically described cells lining in oviduct wall and contributes to interstitial fibrosis and fertility problems. Telocytes (TCs), a newly discovered peculiar type of stromal cells, have been identified in many organs, including oviduct, with proposed multiple potential bio‐functions. However, with recent increasing reports regarding TCs alterations in disease‐affected tissues, there is still lack of evidence about TCs involvement in AS‐affected oviduct tissues and potential pathophysiological roles. We presently identified normal TCs by their characteristic ultrastructural features and immunophenotype. However, in AS‐affected oviduct tissues, TCs displayed multiple ultrastructural damage both in cellular body and prolongations, with obvious loss of TCs and development of tissue fibrosis. Furthermore, TCs lose their interstitial 3‐D network connected by homocellular or heterocellular junctions between TCs and adjacent cells. And especially, TCs connected to the activated immunocytes (mononuclear cells, eosinophils) and affected local immune state (repression or activation). Meanwhile, massive neutrophils infiltration and overproduced Inducible Nitric Oxide Synthase (iNOS), COX‐2, suggested mechanism of inflammatory‐induced TCs damage. Consequently, TCs damage might contribute to AS‐induced structural and reproductive functional abnormalities of oviduct, probably via: (i) substances, energy and functional insufficiency, presumably, e.g. TC‐specific genetic material profiles, ion channels, cytoskeletal elements, Tps dynamics, etc., (ii) impaired TCs‐mediated multicellular signalling, such as homeostasis/angiogenesis, tissue repair/regeneration, neurotransmission, (iii) derangement of 3‐D network and impaired mechanical support for TCs‐mediated multicellular signals within the stromal compartment, consequently induced interstitial fibrosis, (iv) involvement in local inflammatory process/ immunoregulation and possibly immune‐mediated early pregnancy failure.     

Telocytes in exercise‐induced cardiac growth

Exercise can induce physiological cardiac growth, which is featured by enlarged cardiomyocyte cell size and formation of new cardiomyocytes. Telocytes (TCs) are a recently identified distinct interstitial cell type, existing in many tissues and organs including heart. TCs have been shown to form a tandem with cardiac stem/progenitor cells in cardiac stem cell niches, participating in cardiac regeneration and repair. Although exercise‐induced cardiac growth has been confirmed as an important way to promote cardiac regeneration and repair, the response of cardiac TCs to exercise is still unclear. In this study, 4 weeks of swimming training was used to induce robust healthy cardiac growth. Exercise can induce an increase in cardiomyocyte cell size and formation of new cardiomyocytes as determined by Wheat Germ Lectin and EdU staining respectively. TCs were identified by three immunofluorescence stainings including double labelling for CD34/vimentin, CD34/platelet‐derived growth factor (PDGF) receptor‐α and CD34/PDGF receptor‐β. We found that cardiac TCs were significantly increased in exercised heart, suggesting that TCs might help control the activity of cardiac stem/progenitor cells, cardiomyocytes or endothelial cells. Adding cardiac TCs might help promote cardiac regeneration and renewal.     

瘤背石磺的形态、习性和生殖行为

2003年5月～2004年5月研究了上海崇明瘤背石磺(Onchidium struma)的形态特征、生活习性,分别对其消化、呼吸、循环、排泄、生殖、神经等六大系统进行了阐述。结果表明,瘤背石磺生活在潮间带高潮区滩涂的芦苇丛里,摄食泥滩上的有机质和单胞藻类;雌雄同体、异体交配、卵生。生殖系统包括生殖器和雌、雄交接器三部分：生殖器由两性腺、卵黄腺和蛋白腺组成;雄性交接器由输精管、附性腺、阴茎、刺激器等组成,雄性生殖孔位于右侧第一触角中部;雌性交接器由输卵管、受精囊、阴道等组成,雌性生殖孔位于肛门右侧约5．0mm处的腹足与外套膜的交界处。本文并详细描述了石磺的交配行为。     

粉尘螨生殖系统超微结构的透射电镜观察

本研究主要采用透射电镜观察粉尘螨Dermatophagoides farinae (Hughes)生殖系统超微结构。粉尘螨雄性生殖系统是由精巢、 输 精管、 附腺、 射精管、 交配器官及附属交配器官组成。精巢内可同时有精子发育各阶段的细胞。精子无核膜、 核染色质聚集成束、 线 粒体缺乏典型的嵴、 胞质内有平行排列的电子致密薄片等为其特征性结构。雌性生殖系统由交合囊、 交合囊管、 储精囊、 囊导管、 卵 巢、 输卵管、 子宫及产卵管构成。卵巢内可见含多个细胞核的中央细胞, 其周为卵母细胞等生殖细胞。该研究丰富了对粉尘螨生殖系统 结构的认识。     

四眼斑水龟雌性生殖器官组织结构的季节变化

应用石蜡常规切片,HE染色,分别在3月（春季）、8月（夏季）、10月（秋季）、1月（冬季）,对24只四眼斑水龟雌性生殖器官组织结构变化进行研究,结果表明：四眼斑水龟雌性生殖器官的形态和结构存在季节性的变化,8月卵泡开始发育,卵巢增重,子宫腺增多;10月卵巢增重达最大,卵泡发育成熟开始排卵,随后形成黄体,进入排卵期。翌年的1～3月卵巢重量减轻,输卵管管腔增大,腺体增多,子宫肌层不断增厚,进入产卵期。     

Genetic comparison of mouse lung telocytes with mesenchymal stem cells and fibroblasts

Telocytes (TCs) are interstitial cells with telopodes – very long prolongations that establish intercellular contacts with various types of cells. Telocytes have been found in many organs and various species and have been characterized ultrastructurally, immunophenotypically and electrophysiologically ( www.telocytes.com ). Telocytes are distributed through organ stroma forming a three‐dimensional network in close contacts with blood vessels, nerve bundles and cells of the local immune system. Moreover, it has been shown that TCs express a broad range of microRNAs, such as pro‐angiogenic and stromal‐specific miRs. In this study, the gene expression profile of murine lung TCs is compared with other differentiated interstitial cells (fibroblasts) and with stromal stem/progenitor cells. More than 2000 and 4000 genes were found up‐ or down‐regulated, respectively, in TCs as compared with either MSCs or fibroblasts. Several components or regulators of the vascular basement membrane are highly expressed in TCs, such as Nidogen, Collagen type IV and Tissue Inhibitor of Metalloproteinase 3 (TIMP3). Given that TCs locate in close vicinity of small vessels and capillaries, the data suggest the implication of TCs in vascular branching. Telocytes express also matrix metalloproteases Mmp3 and Mmp10, and thus could regulate extracellular matrix during vascular branching and de novo vessel formation. In conclusion, our data show that TCs are not fibroblasts, as the ultrastructure, immunocytochemistry and microRNA assay previously indicated. Gene expression profile demonstrates that TCs are functionally distinct interstitial cells with specific roles in cell signalling, tissue remodelling and angiogenesis.     

体外培养日本血吸虫成虫生殖器官超微结构的观察

将日本血吸虫成虫于851培养基中培养23天后,对其生殖器官进行透射电镜观察。观察结果显示,雌虫卵巢内卵母细胞出现不同程度的变性、坏死;成熟卵黄细胞的卵黄小滴融合,脂质小滴数量增多、体积增大;培养后期卵壳形成发生障碍,最终导致无活性、无卵壳的畸形卵形成。超微结构观察首次显示,体外初产期虫卵卵壳中有条带状低电子密度区和高电子密度区相间排列。     

Cardiac telocytes — their junctions and functional implications

Telocytes (TCs) form a cardiac network of interstitial cells. Our previous studies have shown that TCs are involved in heterocellular contacts with cardiomyocytes and cardiac stem/progenitor cells. In addition, TCs frequently establish 'stromal synapses' with several types of immunoreactive cells in various organs ( www.telocytes.com ). Using electron microscopy (EM) and electron microscope tomography (ET), we further investigated the interstitial cell network of TCs and found that TCs form 'atypical' junctions with virtually all types of cells in the human heart. EM and ET showed different junction types connecting TCs in a network (puncta adhaerentia minima, processus adhaerentes and manubria adhaerentia). The connections between TCs and cardiomyocytes are 'dot' junctions with nanocontacts or asymmetric junctions. Junctions between stem cells and TCs are either 'stromal synapses' or adhaerens junctions. An unexpected finding was that TCs have direct cell-cell (nano)contacts with Schwann cells, endothelial cells and pericytes. Therefore, ultrastructural analysis proved that the cardiac TC network could integrate the overall 'information' from vascular system (endothelial cells and pericytes), nervous system (Schwann cells), immune system (macrophages, mast cells), interstitium (fibroblasts, extracellular matrix), stem cells/progenitors and working cardiomyocytes. Generally, heterocellular contacts occur by means of minute junctions (point contacts, nanocontacts and planar contacts) and the mean intermembrane distance is within the macromolecular interaction range (10-30 nm). In conclusion, TCs make a network in the myocardial interstitium, which is involved in the long-distance intercellular signaling coordination. This integrated interstitial system appears to be composed of large homotropic zones (TC-TC junctions) and limited (distinct) heterotropic zones (heterocellular junctions of TCs).     

Telocytes in the human kidney cortex

Renal interstitial cells play an important role in the physiology and pathology of the kidneys. As a novel type of interstitial cell, telocytes (TCs) have been described in various tissues and organs, including the heart, lung, skeletal muscle, urinary tract, etc. ( www.telocytes.com ). However, it is not known if TCs are present in the kidney interstitium. We demonstrated the presence of TCs in human kidney cortex interstitium using primary cell culture, transmission electron microscopy (TEM) and in situ immunohistochemistry (IHC). Renal TCs were positive for CD34, CD117 and vimentin. They were localized in the kidney cortex interstitial compartment, partially covering the tubules and vascular walls. Morphologically, renal TCs resemble TCs described in other organs, with very long telopodes (Tps) composed of thin segments (podomers) and dilated segments (podoms). However, their possible roles (beyond intercellular signalling) as well as their specific phenotype in the kidney remain to be established.     

Immunohistochemical characterization and functional identification of mammary gland telocytes in the self‐assembly of reconstituted breast cancer tissue in vitro

Telocyte (TC) as a special stromal cell exists in mammary gland and might play an important role in the balance of epithelium‐stroma of mammary gland. Considering that different types of breast interstitial cells influence the development and progression of breast cancer, TCs may have its distinct role in this process. We here studied the roles of TCs in the self‐assembly of reconstituted breast cancer tissue. We co‐cultured primary isolated TCs and other breast stromal cells with breast cancer EMT‐6 cells in collagen/Matrigel scaffolds to reconstitute breast cancer tissue in vitro. Using histology methods, we investigated the immunohistochemical characteristics and potential functions of TCs in reconstituted breast cancer tissue. TCs in primary mammary gland stromal cells with long and thin overlapping cytoplasmic processes, expressed c‐kit/CD117, CD34 and vimentin in reconstitute breast cancer tissue. The transmission electron microscopy showed that the telocyte‐like cells closely communicated with breast cancer cells as well as other stromal cells, and might serve as a bridge that directly linked the adjacent cells through membrane‐to‐membrane contact. Compared with cancer tissue sheets of EMT‐6 alone, PCNA proliferation index analysis and TUNEL assay showed that TCs and other breast stromal cells facilitated the formation of typical nest structure, promoted the proliferation of breast cancer cells, and inhibited their apoptosis. In conclusion, we successfully reconstituted breast cancer tissue in vitro, and it seems to be attractive that TCs had potential functions in self‐assembly of EMT‐6/stromal cells reconstituted breast cancer tissue.     

Telocytes in human heart valves

Valve interstitial cells (VICs) are responsible for maintaining the structural integrity and dynamic behaviour of the valve. Telocytes (TCs), a peculiar type of interstitial cells, have been recently identified by Popescu's group in epicardium, myocardium and endocardium (visit www.telocytes.com ). The presence of TCs has been identified in atria, ventricles and many other tissues and organ, but not yet in heart valves. We used transmission electron microscopy and immunofluorescence methods (double labelling for CD34 and c‐kit, or vimentin, or PDGF Receptor‐β) to provide evidence for the existence of TCs in human heart valves, including mitral valve, tricuspid valve and aortic valve. TCs are found in both apex and base of heart valves, with a similar density of 27–28 cells/mm² in mitral valve, tricuspid valve and aortic valve. Since TCs are known for the participation in regeneration or repair biological processes, it remains to be determined how TCs contributes to the valve attempts to re‐establish normal structure and function following injury, especially a complex junction was found between TCs and a putative stem (progenitor) cell.     

Comparison of Chromosome 4 gene expression profile between lung telocytes and other local cell types

Telocytes (TCs) are new cellular entities of mesenchymal origin described almost ubiquitously in human and mammalian organs ( www.telocytes.com ). Different subtypes of TCs were described, all forming networks in the interstitial space by homo‐ and heterocellular junctions. Previous studies analysed the gene expression profiles of chromosomes 1, 2, 3, 17 and 18 of murine pulmonary TCs. In this study, we analysed by bioinformatics tools the gene expression profiles of chromosome 4 for murine pulmonary TCs and compared it with mesenchymal stem cells (MSCs), fibroblasts (Fbs), alveolar type II cells (ATII), airway basal cells, proximal airway cells, CD8(+) T cells from bronchial lymph nodes (T‐BL) and CD8(+) T cells from lungs (T‐L). Key functional genes were identified with the aid of the reference library of the National Center for Biotechnology Information Gene Expression Omnibus database. Seventeen genes were up‐regulated and 56 genes were down‐regulated in chromosome 4 of TCs compared with other cells. Four genes (Akap2, Gpr153, Sdc3 and Tbc1d2) were up‐regulated between one and fourfold and one gene, Svep1, was overexpressed over fourfold. The main functional networks were identified and analysed, pointing out to a TCs involvement in cellular signalling, regulation of tissue inflammation and cell expansion and movement.     

Female reproductive biology of Platygaster diplosisae (Hymenoptera: Platygastridae) and Aprostocetus procerae (Hymenoptera: Eulophidae), two parasitoids associated with the African Rice Gall Midge, Orseolia oryzivora (Diptera: Cecidomyiidae)

We investigated the female reproductive system of Platygaster diplosisae (Hymenoptera: Platygastridae) and Aprostocetus procerae (= Tetrastichus pachydiplosisae) (Hymenoptera: Eulophidae), two parasitoids associated with the African rice gall midge, Orseolia oryzivora (Diptera: Cecidomyiidae). Both optical and electron microscopy were used. The female reproductive system of P. diplosisae includes two large ovaries of the meristic polytrophic‐type, each composed of several tens of ovarioles. The system includes also a venomous gland that extends to a common oviduct. This gland had a filiform secretory portion, in which the epithelium was thin and surrounded a common evacuation canal. The secretory cells secrete into a large reservoir. Parasitism due to P. diplosisae is gregarious. The female reproductive system of A. procerae includes two ovaries of the meristic polytrophic‐type, and each ovary has a few ovarioles. Each ovariole includes one or two oocytes, which can be seen in the vitellarium. Two accessory glands, which extend to the oviduct, are also visible. The secretory epithelium of the accessory gland is made up of a dense network of secretory cells surrounded by muscle fibers. Females of A. procerae pierce the tissues of the gall and probably deposit one egg on or close to the pupa of the midge. Aprostocetus procerae is a solitary parasitoid of the midge. The two parasitoids exploit the same host at different developmental stages. These findings improve our knowledge of the reproductive biology of these two parasitoids associated with the African rice gall midge, an important pest in Africa.