Basal Lamina Formation by Epidermal Cells in Cell Culture

Author(s):  
Takae Hirone ◽  
Shigeru Taniguchi
1997 ◽  
Vol 5 (3) ◽  
pp. 153-156 ◽  
Author(s):  
Andre Bahoric ◽  
A Robertson Harrop ◽  
Howard M Clarke ◽  
Ronald M Zuker

This in vitro study investigated the possibility of delivering epidermal cells to cell culture plates using a simple and inexpensive aerosolization apparatus. Full-thickness skin biopsies were incubated in dispase to separate the epidermis from the dermis, and the epidermis was treated with trypsin to separate the epidermal cells from one another. The cells were suspended in aerosolization medium (1x106 cells/mL) and sprayed onto culture plates using a sterilized pump-action aerosol nozzle. The plates were incubated and microscopically examined on a daily basis. The aerosolization process was successful in consistently delivering a uniform distribution of suspended epidermal cells. By day 4 there was evidence of cell proliferation, and by days 7 to 9 a confluent layer of cells was achieved on the plates. The monolayer consisted primarily of keratinocytes interspersed with a few fibroblasts. The aerosol method was shown to be effective at delivering a suspension of viable epidermal cells to a culture plate.


Development ◽  
1983 ◽  
Vol 75 (1) ◽  
pp. 49-66
Author(s):  
J. S. H. Taylor ◽  
Alan Roberts

We have described the development of the primary sensory system of the trunk region of Xenopus laevis embryos from larval stages 21 to 32. The system is based upon Rohon-Beard and extramedullary cells, which have central axons forming a dorsolateral spinal tract and peripheral neurites which innervate the skin. The pioneer axons of the central tract grow along the outer surface of the cord at stage 22. These pioneer axons may be used by secondary axons as a growth substrate. As the tract forms it is covered by the radially expanding distal processes, ‘end feet’, of the ependymal cells of the cord. Cell bodies of the extramedullary cells bulge out of the cord surface, and are first seen between the newly segmented myotomes, at stage 24. Peripheral neurites from these extramedullary cells grow out laterally from the cord. The Rohon-Beard cells, located within the cord, produce similar peripheral neurites which grow laterally with the extramedullary cell neurites, using them as a substrate. The neurites form bundles which coincide with the intermyotomes and are periodically spaced. The growth cones of these neurites contact the outer surface of the myotomes and proceed ventrally, first on the myotomes and then on the basal lamina of the skin. ‘Pioneer’ neurites are used by later neurites as a growth substrate, but not to the exclusion of all other substrates. The neurites form a plexus on the skin's basal lamina and contact the underlying epidermal cells through holes in the basal lamina. These holes occur in positions over the intercellular boundaries of the epidermal cells.


1978 ◽  
Vol 79 (2) ◽  
pp. 356-370 ◽  
Author(s):  
C L Marcelo ◽  
Y G Kim ◽  
J L Kaine ◽  
J J Voorhees

A population of neonatal mouse keratinocytes (epidermal basal cells) was obtained by gentle, short-term trypsin separation of the epidermal and dermal skin compartments and discontinuous Ficoll gradient purification of the resulting epidermal cells. Over 4--6 wk of culture growth at 32--33 degrees C, the primary cultures formed a complete monolayer that exhibited entire culture stratification and upper cell layer shedding. Transmission and scanning electron microscopy demonstrated that the keratinocyte cultures progressed from one to two cell layers through a series of stratification and specialization phenomena to a six to eight cell layer culture containing structures characteristic of epidermal cells and resembling in vivo epidermal development. The temporal development of primary epidermal cell culture specialization was confirmed by use of two histological techniques which differentially stain the specializing upper cell layers of neonatal mouse skin. No detectable dermal fibroblast co-cultivation was demonstrated by use of the leucine aminopeptidase histochemical technique and routine electron microscope surveillance of the cultures. Incorporation of [3H]thymidine ([3H]Tdr) was greater than 85% into DNA and was inhibited by both 20 micron cytosine arabinoside (Ara-C) and low temperature. Autoradiography and 90% inhibition of [3H]Tdr incorporation by 2 mM hydroxyurea indicated that keratinocyte culture DNA synthesis was scheduled (not a repair phenomenon). The primary keratinocytes showed an oscillating pattern of [3H]Tdr incorporation into DNA over the initial 23--25 days of growth. Autoradiography demonstrated that the cultures contained 10--30% proliferative stem cells from days 2-25 of culture. The reproducibility of both the proliferation and specialization patterns of the described primary epidermal cell culture system indicates that these cultures are a useful tool for investigations of functioning epidermal cell homeostatic control mechanisms.


Parasitology ◽  
1970 ◽  
Vol 61 (2) ◽  
pp. 177-190 ◽  
Author(s):  
V. R. Southgate

The relationship between the ciliated epidermal cells and the subepidermal layer of the miracidium of Fasciola hepatica has been described. Non-ciliated ridge-like extensions of the subepidermal layer separate the ciliated epidermal cells from each other. The sunken portions of the subepidermal layer, each containing a nucleus, lie below the outer body wall muscles of the miracidium and open into the ridge by narrow neck-like connexions. Elongate vesicles, which may be a source of stored plasma membrane similar to that which occurs in the transitional epithelium of other animals, fill most of the ridge. In addition, characteristic round electron dense granules are found in the ridge but the majority are found in the sunken portions of the subepidermal layer.The development and origins of the tegument of the sporocyst of F. hepatica have been described at the ultrastructural level. When the miracidium is in the process of penetrating the snail host, large vacuoles appear between the ciliated epidermal cells and the basal lamina which overlies the muscles of the body wall. These vacuoles have the effect of loosening the epidermal cells from the basal lamina of the body wall of the miracidium. Possible mechanisms involved in the formation of such vacuoles are suggested and discussed.During penetration of the snail the ciliated epidermal cells of the miracidium are lost; the ridge, a syncytial layer between the epidermal cells which is connected with the subepidermal layer, spreads over the basal lamina and exposed body wall muscles of the metamorphosing sporocyst to form the new outer covering of the sporocyst.Cytoplasm passes from the subtegumentary layer into the tegument during this stage of the development of the body wall of the sporocyst. Muscular contraction and microtubules may be involved in the outward movements of this cytoplasm. The nuclei of the subtegumentary layer remain below the muscles of the body wall.Twenty-four hours after penetration of the snail the outer plasma membrane of the tegument forms folds, which greatly increase the surface area.Sixty hours after penetration involutions between the folds, which may indicate pinocytosis, are present, and it is suggested that pinocytosis may play a role in food absorption.The fully formed tegument is a syncytial layer containing numerous electron dense granules, vacuoles, mitochondria and lipid droplets.The results on the formation of the tegument of the sporocyst have been discussed with reference to the controversy about the origins and terminology of the outer covering of the Platyhelminths.


2012 ◽  
Vol 427 ◽  
pp. 55-59
Author(s):  
Dong Mei Zhang ◽  
Jin Sheng Liang ◽  
Yan Ding ◽  
Xi Mu ◽  
Lei Wang

The effect of activated water on the growth of epidermal cells of newborn mice is studied by cell culture in vitro. To investigate the influence of activated water on the growth of cells, parameters of water are measured. Then, biological mechanism about how the water effects on cells is discussed. It shows that activated water has the promoter action to growth of cells comparing with non-activated water by detecting optical density (OD) at 570 nm with the MTT colorimetry. Statistically, the probability P, less than 0.01, is obvious. We believe that water cluster has become smaller and small water clusters are beneficial for transporting nutrients.


1971 ◽  
Vol 49 (2) ◽  
pp. 247-263 ◽  
Author(s):  
Walter S. Krawczyk

Epidermal repair during wound healing is under investigation at both the light and electron microscopic levels. Suction-induced subepidermal blisters have been employed to produce two complementary model wound healing systems. These two model systems are: (a) intact subepidermal blisters, and (b) opened subepidermal blisters (the blister roof was removed immediately after induction, leaving an open wound). From these studies a pattern of movement for epidermal cells in wound healing is proposed. This pattern of movement is the same for both model systems. Epidermal cells appear to move by rolling or sliding over one another. Fine fibers oriented in the cortical cytoplasm may play an important role in the movement of these epidermal cells. Also instrumental in mediating this movement are intercellular junctions (desmosomes) and a firm attachment to a substrate through hemidesmosomes. In the intact subepidermal blisters hemidesmosomal attachment is made to a continuous and homogeneous substrate, the retained basal lamina. In the opened subepidermal blisters contact of epidermal cells is made to a discontinuous substrate composed of sporadic areas of fibrin and underlying mesenchymal cells.


1984 ◽  
Vol 72 (1) ◽  
pp. 163-172 ◽  
Author(s):  
J. Ellison ◽  
D.R. Garrod

An electron microscopical study of the epidermal-dermal junction in the axolotl and adult Rana pipiens has been carried out. This shows that filaments of about 12nm in diameter, known as anchoring filaments, pass from the hemidesmosomes at the base of the epidermal cells across the basal lamina to the dermis. There they may unite to form broader fibres, known as anchoring fibrils, or may simply form bundles. In the axolotl, particularly, the anchoring fibrils or bundles of anchoring filaments, enmesh with the collagen fibres of the dermis. Removal of epidermal cells with EDTA results in separation along a plane in the lamina rara of the basal lamina, i.e. between the plasma membrane of the cells and the lamina densa. The anchoring filaments remain inserted into the lamina densa. Hemidesmosomal plaques are no longer visible in regions of the plasma membrane that have been separated from the basal lamina by EDTA, and no evidence was found that plaques are engulfed by the cells. It is proposed that the hemidesmosome-anchoring filament system provides a structural link between the collagenous filament system of the dermis and the intracellular cytokeratin filament system of the epidermis, which, in turn, is linked between cells by desmosomes.


1996 ◽  
Vol 74 (12) ◽  
pp. 2143-2153 ◽  
Author(s):  
Karsten Meyer ◽  
Thomas Bartolomaeus

Several members of the Annelida bear apically curved or hooked setae that are aligned in a transverse row inside the neuropodial rim. Based on the hypothesis that these specific setae characterize a monophyletic group within the Annelida, the structure and development of the hooked seta in Owenia fusiformis are analysed and compared with data from other annelids with such setae. The neuropodial hooks of O. fusiformis are arranged in multiple transversal rows or setal patches on each side of the body from the fourth setiger onwards. The setae are curved distally and consist of two identical spines lying side by side at the same level. Their tips generally point ventrofrontally. Within each patch, the setae lie inside a setal follicle that consists of a basal chaetoblast, at least one follicle cell, and varying numbers of epidermal cells. Each setal patch is basally surrounded by an extracellular matrix that is continuous with the subepidermal basal lamina. An additional discontinuous extracellular matrix lies between the epidermis and the follicle cells. It is of functional significance for the attachment of the epidermal cells and seems to be related to the special organization of the setal patches, because it is absent in juveniles; they have single neuropodial rows of hooked setae per segment. New setae are formed at the dorsal and caudal edges of each patch, whereas the degeneration of setae is observed at the frontal edge of each patch. Microvilli project from the apex of the chaetoblast into canals within the fully differentiated setae. These canals remain when the microvilli are withdrawn from the seta during formation. Each hook is formed by a single large microvillus. The results of the present paper substantiate the hypothesis of a homology of the hooked setae in the Oweniida and other Annelida. These results and data from the literature support the hypothesis that the Oweniida is the sister-group of a monophylum which consists of the Terebellida, Pogonophora, and Sabellida.


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