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1.
Activation of epithelial stem cells and efficient recruitment of their proliferating progeny plays a critical role in cutaneous wound healing. The reepithelialized wound epidermis has a mosaic composition consisting of progeny that can be traced back both to epidermal and several types of hair follicle stem cells. The contribution of hair follicle stem cells to wound epidermis is particularly intriguing as it involves lineage identity change from follicular to epidermal. Studies from our laboratory show that hair follicle-fated bulge stem cells commit only transient amplifying epidermal progeny that participate in the initial wound re-epithelialization, but eventually are outcompeted by other epidermal clones and largely disappear after a few months. Conversely, recently described stem cell populations residing in the isthmus portion of hair follicle contribute long-lasting progeny toward wound epidermis and, arguably, give rise to new interfollicular epidermal stem cells. The role of epithelial stem cells during wound healing is not limited to regenerating stratified epidermis. By studying regenerative response in large cutaneous wounds, our laboratory uncovered that epithelial cells in the center of the wound can acquire greater morphogenetic plasticity and, together with the underlying wound dermis, can engage in an embryonic-like process of hair follicle neogenesis. Future studies should uncover the cellular and signaling basis of this remarkable adult wound regeneration phenomenon.  相似文献   

2.
A morphological study of in vitro wound healing has been performed by light, transmission and scanning electron microscopy in dorsal thoraco-lumbar skin of 7-day chick embryos. A circular wound, 750 microns in diameter, was punched out of dorsal skin, removing epidermis and the underlying dense dermis. Wound closure was completed within 96 to 120 hours. Feather bud development was not observed at the wound site. The epidermis began to migrate some 24 h after the wounding; the migration of peridermal cells preceded that of basal epidermal cells by some 12 hours. Mechanisms of the epidermal migration were similar to those observed in situ during wound healing of the integument in 5-day chick embryos (THEVENET, 1981), Superficial epithelization of bare dermis occurred as soon as 12 h after the injury. Cytoplasm of dermal cells exhibited many microtubules and a dilated rough endoplasmic reticulum. During the first 48 h, the epidermal cells established direct contacts and zones of close parallel apposition with epithelized dermal cell processes. The basement membrane lamina densa was maintained at the edges of the wound without retraction or ruffling. It was reconstituted concomitantly with the epidermal migration within 72 h. Cytoplasm of migratory epidermal and epithelized dermal cells exhibited many cytoskeleton structures.  相似文献   

3.
Incisional wounds of the same length and depth were made in the skin of catfish Heteropneustes fossilis , and the response in terms of levels of glycogen in cells during healing was studied histochemically. After injury, glycogen disappeared completely from the cells in the migrating epidermis. This disappearance is correlated with the migration of the epidermal cells toward the wound gap for epithelialization and the increased production of mucous cells at the initial stages of healing. The gradual accumulation of glycogen in the epithelial cells and the basal cells after epithelialization of the wound during the early stages of healing suggests that in these cells gluconeogenesis predominates. The gradual disappearance of glycogen from the epithelial cells 72 h after injury and from the basal cells 36 h after injury indicates the gradual resumption of their metabolic activities. The increased gradients of glycogen in club cells that are undergoing vacuolization and disintegration are discussed. No significant levels of glycogen were observed during the formation of granulation tissue and biosynthesis of the sub-cutis. The depletion of glycogen in the degenerating muscle bundles is correlated with the synthesis of lactic acid which contributes to local acidity and favours the autolysis of muscles.  相似文献   

4.
The discovery of long-lived epithelial stem cells in the bulge region of the hair follicle led to the hypothesis that epidermal renewal and epidermal repair after wounding both depend on these cells. To determine whether bulge cells are necessary for epidermal renewal, here we have ablated these cells by targeting them with a suicide gene encoding herpes simplex virus thymidine kinase (HSV-TK) using a Keratin 1-15 (Krt1-15) promoter. We show that ablation leads to complete loss of hair follicles but survival of the epidermis. Through fate-mapping experiments, we find that stem cells in the hair follicle bulge do not normally contribute cells to the epidermis which is organized into epidermal proliferative units, as previously predicted. After epidermal injury, however, cells from the bulge are recruited into the epidermis and migrate in a linear manner toward the center of the wound, ultimately forming a marked radial pattern. Notably, although the bulge-derived cells acquire an epidermal phenotype, most are eliminated from the epidermis over several weeks, indicating that bulge stem cells respond rapidly to epidermal wounding by generating short-lived 'transient amplifying' cells responsible for acute wound repair. Our findings have implications for both gene therapy and developing treatments for wounds because it will be necessary to consider epidermal and hair follicle stem cells as distinct populations.  相似文献   

5.
Hamster tracheal epithelia consist of three cell types: ciliated, mucus and basal cells. Autoradiographic data from several studies suggest that either basal or non-ciliated columnar cells may serve as stem cells for regeneration of lost or damaged ciliated and mucus cells. The objective of the present study was to examine the role of basal cells in the formation of ciliated and mucus cells in hamster tracheal epithelial (HTE) cell cultures via tritiated thymidine ([3H]-TdR) autoradiography. When 3 day cultures were pulsed with [3H]-TdR for 6 hr and incubated for 2 additional days in non-radioactive media (5 day total) label was present in the nuclei of basal and columnar epithelial cells suggesting that the labeled columnar cells may be derived from basal cells. However, the morphological reorganization occurring during this 2 day interval may create difficulties in this interpretation. Since these morphological changes are minimal during the 6 day to 8 day in vitro period, 6 day HTE cultures were pulsed with [3H]-TdR for 6 hr and incubated for 2 additional days in non-radioactive media (8 day total), and examined to further study the fate of labeled basal cells during this period. Analysis of these 8 day cultures revealed that labeled nuclei were present in both basal cells and adjacent ciliated and mucus cells. These results do not exclude the possibility of non-basal cell origin of ciliated and mucus cells in other systems but suggest that, at least in HTE cultures, undifferentiated basal cells have the ability to develop into ciliated and mucus cells.  相似文献   

6.
The epidermis of Geocentrophora wagini was studied using transmission electron microscopy. The turbellarian body was entirely covered by cilia, whose density was higher on the ventral surface compared with the dorsal one. In all regions examined, the epidermis was made up of a one-layered insunk epithelium. The basal matrix, underlying the epidermis, was a well developed basement membrane (BM) with bilayered structure, overlying the muscle network of circular and longitudinal fibers. The double plasma membranes, extending from the apical surface of epidermis to BM, were linked by specialized cell junctions. This suggested that epidermis had a cellular rather than a cyncytial arrangement. Each insunk epidermal cell was made of two unequal parts: a comparatively thin surface plate attached to BM by hemiadherens junctions, and a massive nucleated portion located below the body wall musculature in the parenchyma. A thin cytoplasmic bridge connected the epidermal plate with the nucleated cell body. The epidermal plates were joined by belt-like junctions along their adjacent surfaces. Inconspicuous zonula adherens (ZA) had a most apical position, and prominent septate junction was arrayed proximally to this zonula. Except ZA, cell boundaries in epidermis were frequently flanked by rows of light tubules and vesicles. In the basal half of the epithelial sheet, they were occassionally accompanied by single cisternae of rough endoplasmic reticulum (RER). The ultrastructure of the insunk cell body and that of the surface plate showed a considerable similarity. The common features were distinctive profiles of RER and GA, the presence of epitheliosomes, light tubules and vesicles, centrioles and fibrous granules. Thus, ultrastructural features allow a rather reliable identification of epidermal cells in the parenchyma, despite the absence of any visible morphological association between cell body and its epidermal plate.  相似文献   

7.
The epidermis serves as a barrier protecting organs and tissues from the environment, and comprises many types of cells. A cell renewal system is established in epidermis: old epithelial cells are replaced by newly differentiated cells, which are derived from epidermal stem cells located near basement membrane. In order to examine the mechanism of epidermal development, we isolated a novel gene expressed in Xenopus epidermis and named the gene Xenopus polka dots (Xpod) from its polka dot-like expression pattern throughout larval periods. Several immunohistochemical examinations showed that the Xpod-expressing cell type is neither p63-positive epidermal stem cells, nor the α-tubulin-positive ciliated cells, but a subset of the foxi1e-positive ionocytes. The forced gene expression of foxi1e caused the suppression of Xpod expression, while Xpod showed no effect on foxi1e expression. In a comparison of several osmotic conditions, we found that hypertonic culture caused the increase in number of the Xpod-expressing cell, whereas number of the foxi1e-expressing cells was reduced under the hypertonic condition. These results show the possibility that Xpod is involved in the establishment of a certain subpopulation of ionocytes under hypertonic conditions.  相似文献   

8.
In this review we focus on epidermal stem cells in the normal regeneration of the skin as well as in wounded and psoriatic skin. Furthermore, we discuss current data supporting the idea of cancer stem cells in the pathogenesis of skin carcinoma and malignant melanoma. Epidermal stem cells present in the basal layer of the interfollicular epidermis and in the bulge region of the hair follicle play a critical role for normal tissue maintenance. In wound healing, multipotent epidermal stem cells contribute to re-epithelization. It is possible that defects in growth control of either epidermal stem cells or transit amplifying cells constitute a primary pathogenetic factor in the epidermal hyperproliferation seen in psoriasis. In cutaneous malignancies mounting evidence supports a stem cell origin in skin carcinoma and malignant melanoma and a possible existence of cancer stem cells.  相似文献   

9.
通过银染法对家蚕整体染色,结果表明:家蚕消化管内的桑叶由叶表皮、叶肉和叶脉组成。叶表皮包括上表皮和下表皮。上表皮细胞可分为三种:钟乳体细胞、绿色表皮细胞和黄色表皮细胞;下表皮内含有气孔;叶肉组织内含有晶体,其中海绵组织内的最多。家蚕消化管由前向后可分为前肠、中肠和后肠,由外向内依次为肌层、底膜、上皮细胞层、内膜。中肠最为发达,其发达的上皮细胞向内表面突起形成许多大的指突形皱褶;上皮细胞层内有圆筒形细胞、杯形细胞两种细胞,两者在形状、功能以及嗜银性等方面有所差异。家蚕消化管对桑叶不同组织的消化吸收效率有差异,上表皮吸收效率最高,下表皮和栅栏组织次之,最低的是海绵组织。采用动物细胞染色方法对植物细胞进行染色,并与常规植物学染色方法进行了比较;依据细胞嗜银性的不同,可将桑叶的上表皮细胞分为两种亚型。  相似文献   

10.
At 10 days in ovo the embryonic chick epidermis acquires the ability to spread as a cohesive epithelial sheet when wounded. A tissue culture system has been constructed that supports epidermal cell outgrowth consistent with epidermal behaviour in vivo and permits experimental manipulation of the isolated tissue with growth factors and other hormones. This culture system consists of embryonic chick epidermis isolated at days 8, 10, and 12 of development, serum-free, chemically-defined culture medium, and the inner surface of the vitelline membrane of the hen's egg as the culture substratum. At 8 days the cellular outgrowth is mesenchymal in the absence of exogenous EGF. The 8 day tissues responds to added EGF by exhibiting precocious epithelial outgrowth. The results suggest that sensitivity to EGF or EGF-like growth factors is part of the mechanism underlying the developmental onset of epidermal wound healing in skin. The epidermal origin of the outgrowth is determined by antibody staining for specific cytokeratins. The epithelial character of the outgrowth is determined by visualizing actin microfilament distribution. The normal epithelial outgrowth shows apical/basal polarization of the sheet except at the edge. From 10 days on, the isolated epidermis exhibits epithelial outgrowth from explants in culture in the absence of exogenous EGF, suggesting endogenous production of an EGF-like factor. Glucocorticoid and mineralocorticoid hormones both produce a reduced amount of epithelial outgrowth. This retardation of the early outgrowth by glucocorticoids and mineralocorticoids could result from a reduced ability of the cut edge of the epidermis to 'disorganize' and assume the unpolarized migratory form required for rapid epidermal wound healing.  相似文献   

11.
A therapeutic approach being investigated for a variety of pathologies is tissue regeneration using a patient's own cells. Such studies have been hampered due to the difficulty in growing epithelial cells for prolonged periods in culture. Replicative senescence due to short telomeres and p16 induced by culture stress work together to inhibit cell growth. Forced expression of telomerase (hTERT) can prevent replicative senescence, and expression of the cell cycle protein cdk4 can sequester p16, thereby immortalizing epithelial cells in culture. In the present study, we used this method to immortalize human bronchial epithelial cells (HBECs) to determine whether immortalized HBECs retain the ability to differentiate normally. HBECs were plated atop contracted collagen gels containing lung fibroblasts. This three-dimensional (3D) tissue model was cultured initially submerged, then raised to the air/liquid interface for up to 28 days. Normal differentiation was assessed by the presence of ciliated cells, goblet (mucin-producing) cells, and basal epithelial cells. Scanning electron microscopic observations revealed both ciliated and non-ciliated cells in these 3D tissues. Histological examination revealed the presence of mucin-producing cells, and immunohistochemistry using antibodies against p63 and keratin 14 showed the presence of basal cells. These results demonstrate that immortalized HBECs retain the capacity to differentiate into each of three cell types: basal, mucin-producing, and columnar ciliated epithelial cells. Such cells will be useful cellular reagents for research in aging, cancer progression, as well as normal bronchial epithelial differentiation and will help progress the use of engineered cells to enhance tissue regeneration.  相似文献   

12.
Continuous renewal of the epidermis and its appendages throughout life depends on the proliferation of a distinct population of cells called stem cells. We have used in situ retrovirus-mediated gene transfer to genetically mark cutaneous epithelial stem cells of adolescent mice, and have followed the fate of the marked progeny after at least 37 epidermal turnovers and five cycles of depilation-induced hair growth. Histological examination of serial sections of labeled pilosebaceous units demonstrated a complex cell lineage. In most instances, labeled cells were confined to one or more follicular compartments or solely to sebaceous glands. Labeled keratinocytes in interfollicular epidermis were confined to distinct columnar units representing epidermal proliferative units. The contribution of hair follicles to the epidermis was limited to a small rim of epidermis at the margin of the follicle, indicating that long term maintenance of interfollicular epidermis was independent of follicle-derived cells. Our results indicate the presence of multiple stem cells in cutaneous epithelium, some with restricted lineages in the absence of major injury.  相似文献   

13.
Wound repair in planarians is mainly characterized by two cell-migratory events involving the epidermis adjacent to the wound and its basement membrane. The first event is the migration of epidermal cells to cover the wound surface; the second one is the migration of newly differentiating replacement epidermal cells from the parenchyma to the epidermis. In addition to these events, migration of fixed parenchymal cells is observed during wound healing. All migrating cells were characterized by the presence of actin, as shown by the results obtained by means of indirect immunolocalization with fluorescent and electron microscopy. Migrating cells were heavily labeled with gold particles, which clustered at the level of cell-matrix and cell-cell contacts.  相似文献   

14.
Dorsal closure in Drosophila embryogenesis involves expansion of the dorsal epidermis, followed by closure of the opposite epidermal edges. This process is driven by contractile force generated by an extraembryonic epithelium covering the yolk syncytium known as the amnioserosa. The secreted signaling molecule Dpp is expressed in the leading edge of the dorsal epidermis and is essential for dorsal closure. We found that the outermost row of amnioserosa cells (termed pAS) maintains a tight basolateral cell-cell adhesion interface with the leading edge of dorsal epidermis throughout the dorsal closure process. pAS was subject to altered cell motility in response to Dpp emanating from the dorsal epidermis, and this response was essential for dorsal closure. alphaPS3 and betaPS integrin subunits accumulated in the interface between pAS and dorsal epidermis, and were both required for dorsal closure. Looking at alphaPS3, type I Dpp receptor, and JNK mutants, we found that pAS cell motility was altered and pAS and dorsal epidermis adhesion failed under the mechanical stress of dorsal closure, suggesting that a Dpp-mediated mechanism connects the squamous pAS to the columnar dorsal epidermis to form a single coherent epithelial layer.  相似文献   

15.
Abstract A small syncytium or a pair of syncytia is present in the dorsal epidermis of the four species studied (Diceratocephala boschmai, Temnocephala dendyi, Craspedella sp., and Achenella sathonota). Unlike the other epidermal syncytia, the posttentacular ones may be deeply stained with silver nitrate. The basal zone of the posttentacular syncytia is occupied by abundant deep invaginations of the basal cell membrane with densely packed mitochondria between them. In one species, A. sathonota, the nuclei containing portions of the posttentacular syncytia are insunk into the parenchyma. Ultrastructurally, the posttentacular syncytia are similar to the excretory bladder (the distal portion of the protonephridial canal) of the same animals and to osmoregulatory epithelial chloride cells known from many animals. These data suggest that ion transport in this syncytium serves for osmoregulation and maintenance of ion balance. Despite extensive studies of the epidermis, no hints of such cells have ever been reported in other platyhelminths. The accessory osmoregulatory organ is probably an adaptation to life in the gill chamber of the crustacean hosts which are characterized by high ability to maintain osmotic and ionic balance in stressful conditions.  相似文献   

16.
The dorsal surface of the holothurioid Holothuria forskali bears several longitudinal rows of modified podia called papillae. Each papilla consists of a conical stem topped by an hemispherical bud. Their gross tissue stratification is the same all along the papilla being made up of four tissue layers, viz. an inner mesothelium, a connective tissue layer, a nerve plexus and an outer epidermis. The latter is differently organized according to whether it belongs to the stem or to the bud. The epidermis of the bud is built up by ciliated cells that intimately contact the nerve plexus and have the classical structure of echinoderm sensory cells. The papillae are thus sensory organs involved in mechanoreception and possibly chemoreception.  相似文献   

17.
When the dorsal and ventral epidermal layers join by first intention during the closure of the wound, the cells of their borders (M-cells) do not meet in the same manner in all sections. In anterior sections the dorsal M-cells attach themselves to the ventral basement membrane, so that only the dorsal epidermis is stretched. In posterior sections the dorsal and the ventral M-cells join by their apical edges without being closely apposed to the wound surface. Only the ventral cells are stretched because of their specific motility. In longitudinal sections the dorsal and the ventral M-cells also join by their apical edges, but since they are closely apposed to the wound surface both epidermal layers are stretched. The stretching is a process equivalent to distalization. The junction between the dorsal and the ventral epidermis is shifted ventrally in the anterior wounds (as in the intact heads) and dorsally in the posterior wounds (as in the intact tails). Some abnormalities of wound closure have been observed at levels where heteromorphic regeneration frequently occurs. These findings are consistent with the hypothesis previously advanced (3) that the modalities of wound closure establish the programme for regeneration.  相似文献   

18.
In order to examine the spatial organisation of stem cells and their progeny in human epidermis, we developed a method for whole-mount epidermal immunofluorescence labelling using high surface beta1 integrin expression as a stem cell marker. We confirmed that there are clusters of high beta1 integrin-expressing cells at the tips of the dermal papillae in epidermis from several body sites, whereas alpha6 integrin expression is more uniform. The majority of actively cycling cells detected by Ki67 or bromodeoxyuridine labelling were found in the beta1 integrin-dull, transit amplifying population and integrin-negative, keratin 10-positive cells left the basal layer exclusively from this compartment. When we examined p53-positive clones in sun-exposed epidermis, we found two types of clone that differed in size and position in a way that was consistent with the founder cell being a stem or transit amplifying cell. The patterning of the basal layer implies that transit amplifying cells migrate over the basement membrane away from the stem cell clusters. In support of this, isolated beta1 integrin-dull keratinocytes were more motile on type IV collagen than beta1 integrin-bright keratinocytes and EGFP-labelled stem cell clones in confluent cultured sheets were compact, whereas transit amplifying clones were dispersed. The combination of whole-mount labelling and lineage marking thus reveals features of epidermal organisation that were previously unrecognised.  相似文献   

19.
20.
The epidermis of the land planarian Bipalium adventitium was examined by light and electron microscopy. In all regions, the epidermis consists of a simple columnar ciliated epithelium associated with a prominent basement membrane. The epithelial cells, possessing abundant microvilli and poorly developed terminal webs, are conjoined laterally at their apical ends by septate junctions. The epidermis of the creeping sole is distinguished from that of adjoining regions by a “insunken” condition of the epithelial cells, a greater number of cilia per cell, and an absence of glandular secretions other than mucus. The insunken cells of the sole possess large glycogen disposits and attributes of metabolically active cells. Unusual intranuclear inclusions of unknown significance are also found in many of the epidermal cells in all regions. The basement membrane lacks distinct layering and consists of fine fibrils displaying a beaded appearance but no obvious cross-banding. Histochemical tests indicate that the fibrils are collagenous. In addition to mucus, secretory material found in nonsole regions includes lamellated granules and rhabdites, both stained intensely by acidic dyes. Rhabdites and the basement membrane also contain disulfide-enriched proteins. In scanning electron micrographs, the sole appears as a faint, longitudinally oriented band extending along the entire length of the animal. In all regions except the sensory border of the head, the microvilli are generally obscured by the densely arranged cilia. The sensory border consists of a row of toothlike papillae and grooves covered almost exclusively by microvilli, small club-shaped structures, and larger spherical protrusions.  相似文献   

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