scholarly journals Stratification, specialization, and proliferation of primary keratinocyte cultures. Evidence of a functioning in vitro epidermal cell system.

1978 ◽  
Vol 79 (2) ◽  
pp. 356-370 ◽  
Author(s):  
C L Marcelo ◽  
Y G Kim ◽  
J L Kaine ◽  
J J Voorhees
Keyword(s):  
Cell Culture ◽  
Epidermal Cell ◽  
Cell Layer ◽  
Primary Cultures ◽  
Mouse Skin ◽  
Epidermal Cells ◽  
Neonatal Mouse ◽  
Histological Techniques ◽  
Cell Layers ◽  
Epidermal Cell Culture

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.

scholarly journals Fluorochrome-coupled lectins reveal distinct cellular domains in human epidermis.

1986 ◽  
Vol 34 (3) ◽  
pp. 307-315 ◽  
Author(s):  
I Virtanen ◽  
A L Kariniemi ◽  
H Holthöfer ◽  
V P Lehto
Keyword(s):  
Endothelial Cells ◽  
Epidermal Cell ◽  
Basal Cell ◽  
Cell Layer ◽  
Lectin Binding ◽  
Epidermal Cells ◽  
Human Epidermis ◽  
Binding Pattern ◽  
Basal Cell Layer ◽  
Cell Layers

The distribution of saccharide moieties in human interfollicular epidermis was studied with fluorochrome-coupled lectins. In frozen sections Concanavalin A (Con A), Lens culinaris agglutinin (LCA), Ricinus communis agglutinin I (RCAI), and wheat germ agglutinin (WGA) stained intensively both dermis and viable epidermal cell layers, whereas peanut agglutinin (PNA) bound only to living epidermal cell layers. Ulex europaeus agglutinin I (UEAI) bound to dermal endothelial cells and upper cell layers of the epidermis but left the basal cell layer unstained. Dolichos biflorus agglutinin (DBA) bound only to basal epidermal cells, whereas both soybean agglutinin (SBA) and Helix pomatia agglutinin (HPA) showed strong binding to the spinous and granular cell layers. On routinely processed paraffin sections, a distinctly different staining pattern was seen with many lectins, and to reveal the binding of some lectins a pretreatment with protease was required. All keratin-positive cells in human epidermal cell suspensions, obtained with the suction blister method, bound PNA, whereas only a fraction of the keratinocytes bound either DBA or UEAI. Such a difference in lectin binding pattern was also seen in epidermal cell cultures both immediately after attachment and in organized cell colonies. This suggests that in addition to basal cells, more differentiated epidermal cells from the spinous cell layer are also able to adhere and spread in culture conditions. Gel electrophoretic analysis of the lectin-binding glycoproteins in detergent extracts of metabolically labeled primary keratinocyte cultures revealed that the lectins recognized both distinct and shared glycoproteins. A much different lectin binding pattern was seen in embryonic human skin: fetal epidermis did not show any binding of DBA, whereas UEAI showed diffuse binding to all cell layers but gave a bright staining of dermal endothelial cells. This was in contrast to staining results obtained with a monoclonal cytokeratin antibody, which showed the presence of a distinct basal cell layer in fetal epidermis also. The results indicate that expression of saccharide moieties in human epidermal keratinocytes is related to the stage of cellular differentiation, different cell layers expressing different terminal saccharide moieties. The results also suggest that the emergence of a mature cell surface glycoconjugate pattern in human epidermis is preceded by the acquisition of cell layer-specific, differential keratin expression.

Non-cell-autonomous function of the Antirrhinum floral homeotic proteins DEFICIENS and GLOBOSA is exerted by their polar cell-to-cell trafficking

Development ◽  
1996 ◽  
Vol 122 (11) ◽  
pp. 3433-3441 ◽  
Author(s):  
M.C. Perbal ◽  
G. Haughn ◽  
H. Saedler ◽  
Z. Schwarz-Sommer
Keyword(s):  
Cell Layer ◽  
Epidermal Cells ◽  
Molecular Techniques ◽  
Cell Trafficking ◽  
Mads Box ◽  
Organ Identity ◽  
Cell Layers ◽  
Periclinal Chimeras ◽  
The Difference ◽  
Cell Autonomy

In Antirrhinum majus, petal and stamen organ identity is controlled by two MADS-box transcription factors, DEFICIENS and GLOBOSA. Mutations in either of these genes result in the replacement of petals by sepaloid organs and stamens by carpelloid organs. Somatically stable def and glo periclinal chimeras, generated by transposon excision events, were used to study the non-cell-autonomous functions of these two MADS-box proteins. Two morphologically distinct types of chimeras were analysed using genetic, morphological and molecular techniques. Restoration of DEF expression in the L1 cell layer results in the reestablishment of DEF and GLO functions in L1-derived cells only; inner layer cells retain their mutant sepaloid features. Nevertheless, this activity is sufficient to allow the expansion of petal lobes, highlighting the role of DEF in the stimulation of cell proliferation and/or cell shape and elongation when expressed in the L1 layer. Establishment of DEF or GLO expression in L2 and L3 cell layers is accompanied by the recovery of petaloid identity of the epidermal cells but it is insufficient to allow petal lobe expansion. We show by in situ immunolocalisation that the non-cell-autonomy is due to direct trafficking of DEF and GLO proteins from the inner layer to the epidermal cells. At least for DEF, this movement appears to be polar since DEF acts cell-autonomously when expressed in the L1 cell layer. Furthermore, the petaloid revertant sectors observed on second whorl mutant organs and the mutant margins of petals of L2L3 chimeras suggest that DEF and GLO intradermal movement is limited. This restriction may reflect the difference in the regulation of primary plasmodesmata connecting cells from the same layer and secondary plasmodesmata connecting cells from different layers. We propose that control of intradermal trafficking of DEF and GLO could play a role in maintaining of the boundaries of their expression domains.

Human Hair Follicle Germinative Epidermal Cell Culture

1993 ◽  
Vol 101 (4) ◽  
pp. 634-638 ◽  
Author(s):  
Amanda J Reynolds ◽  
Clifford M Lawrence ◽  
Colin A B Jahoda
Keyword(s):  
Cell Culture ◽  
Hair Follicle ◽  
Epidermal Cell ◽  
Human Hair ◽  
Human Hair Follicle ◽  
Epidermal Cell Culture

Surface modification of oriented polysaccharide scaffolds using biotic nanofibers for epidermal cell culture

Cellulose ◽  
2019 ◽  
Vol 26 (13-14) ◽  
pp. 7971-7981
Author(s):  
Suresh N. Rao ◽  
Yutaro Tsujita ◽  
Tetsuo Kondo
Keyword(s):  
Cell Culture ◽  
Surface Modification ◽  
Epidermal Cell ◽  
Epidermal Cell Culture

scholarly journals Pigment Distribution and Epidermal Cell Shape in Dendrobium Species and Hybrids

HortScience ◽  
2003 ◽  
Vol 38 (4) ◽  
pp. 573-577 ◽  
Author(s):  
Rasika G. Mudalige ◽  
Adelheid R. Kuehnle ◽  
Teresita D. Amore
Keyword(s):  
Epidermal Cell ◽  
Cell Shape ◽  
Epidermal Cells ◽  
Vascular Bundles ◽  
Color Intensity ◽  
Visual Texture ◽  
Intensity Perception ◽  
Cell Layers ◽  
Cell Shapes ◽  
Pigment Distribution

Perianths of 34 Dendrobium Sw. species and hybrids were examined to elucidate the roles of pigment distribution and shape of upper epidermal cells in determining color intensity, perception, and visual texture. Color intensity was determined by the spatial localization of anthocyanin in tissue layers, i.e., in the epidermal, subepidermal, and mesophyll layers, as well as by distribution of pigmented cells within the tissue layer. Anthocyanins were confined to the epidermal layer or subepidermal layer in flowers with low color intensity, whereas they were also in several layers of mesophyll in more intensely colored flowers. Striped patterns on the perianth were due to the restriction of pigment to cells surrounding the vascular bundles. Color perception is influenced by the presence or absence of carotenoids, which when present, were distributed in all cell layers. Anthocyanins in combination with carotenoids resulted in a variety of flower colors ranging from red, maroon, bronze to brown, depending on the relative location of the two pigments. Four types of epidermal cell shapes were identified in Dendrobium flowers: flat, dome, elongated dome, and papillate. Epidermal cell shape and cell packing in the mesophyll affected the visual texture. Petals and sepals with flat cells and a tightly packed mesophyll had a glossy texture, whereas dome cells and loosely packed mesophyll contributed a velvety texture. The labella in the majority of flowers examined had a complex epidermis with more than one epidermal cell shape, predominantly papillate epidermal cells.

Cytoskeleton and pericellular matrix organization of pure adult human keratinocytes cultured from suction-blister roof epidermis

1982 ◽  
Vol 58 (1) ◽  
pp. 49-61
Author(s):  
A.L. Kariniemi ◽  
V.P. Lehto ◽  
T. Vartio ◽  
I. Virtanen
Keyword(s):  
Epidermal Cell ◽  
Cell Cultures ◽  
Primary Cultures ◽  
Epidermal Cells ◽  
Collagen Type Iv ◽  
Direct Role ◽  
Adult Human ◽  
Type Iv ◽  
Suction Blister ◽  
Marginal Cells

Pure adult human keratinocyte cultures were raised from suction-blister roof epidermis and cultured in MCDB-151 medium. In primary culture the epidermal cells rapidly adhered, spread and began to proliferate on collagen-coated growth substrata but not on uncoated plastic or glass substrata. A fibrillar keratin-specific fluorescence, showing a typical cell-cell arrangement, was seen in all cells in indirect immunofluorescence microscopy, whereas only some cells also showed vimentin-specific staining. A fine fibrillar fibronectin-specific surface staining was seen at the margin of attaching cells and in marginal cells of spreading cell islands, whereas no fluorescence could be seen in epidermal cells, with antibodies against type IV collagen or laminin. Interestingly, the marginal cells also showed intracellular fibronectin. The synthesis of fibronectin in epidermal cell cultures could also be revealed by metabolic labelling experiments with [35S]methionine. In contrast to primary cultures, subcultivated keratinocytes also adhered to uncoated plastic and glass substrata. After subcultivation, keratin and surface fibronectin distribution remained unaltered but after some subcultivations, most of the cells also showed fibrillar vimentin and expressed fibronectin intracellularly. The results show that the suction-blister method provides an easy way to obtain pure epidermal cell cultures without contaminating mesenchymal cells. Our results also suggest a direct role for fibronectin but not for collagen type IV or laminin in adhesion and spreading of epidermal cells in vitro.

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