The Use of Cultured Human Keratinocytes as a Model System for Skin Pharmacology and Toxicology

1990 ◽  
pp. 181-191
Author(s):  
Uwe Reichert ◽  
Rainer Schmidt
2014 ◽  
Vol 23 (6) ◽  
pp. 443-446 ◽  
Author(s):  
Rajesh L. Thangapazham ◽  
Peter Klover ◽  
Shaowei Li ◽  
Ji-an Wang ◽  
Leonard Sperling ◽  
...  

1998 ◽  
Vol 72 (1) ◽  
pp. 749-757 ◽  
Author(s):  
Renske D. M. Steenbergen ◽  
Jacqueline N. Parker ◽  
Sharon Isern ◽  
Peter J. F. Snijders ◽  
Jan M. M. Walboomers ◽  
...  

ABSTRACT Organotypic cultures of human keratinocytes provide a useful model system to study human papillomavirus (HPV)-host cell interactions. In this study, we analyzed organotypic cultures of two HPV type 16 (HPV16) (FK16A and FK16B)- and two HPV18 (FK18A and FK18B)-transfected keratinocyte cell lines through the process of immortalization in vitro. For FK16A and FK18B cells, passages of both mortal cells in their extended life span and subsequent immortal stages were studied. Mortal cells of FK16A and FK18B showed a morphology reminiscent of mild to moderate dysplasia, whereas in their immortal descendants, severely dysplastic features were observed. Immortal FK18A cells were mildly to moderately dysplastic, while FK16B cells were severely dysplastic. The increasing degrees of dysplasia were associated with a decreasing expression of differentiation markers cytokeratin 10 and profilaggrin. All raft cultures expressed E6-E7 mRNAs in the basal layer, while the amount of viral transcripts in the suprabasal cells was in general proportional to the degree of dysplasia. In all cases, E6-E7 transcription and dysplastic features were highly correlated with cellular proliferation, as assessed by Ki-67 (MIB-1) antigen expression. Moreover, high levels of E6-E7 transcription and expression of p21cip1 protein in the basal layer seemed to be mutually exclusive. We conclude that expression of E6-E7 in the basal cells associated with increased proliferation in the absence of detectable p21cip1 protein is apparently necessary but not sufficient for immortalization, or for the loss of terminal differentiation, for which yet to be discovered additional events are required. The model system described in this study provides a valuable tool to analyze alterations in viral transcription regulation during HPV-mediated cell transformation.


Author(s):  
D. C. Williams ◽  
D. E. Outka

Many studies have shown that the Golgi apparatus is involved in a variety of synthetic activities, and probably no Golgi product is more elaborate than the scales produced by various kinds of phytoflagellates. The formation of calcified scales (coccoliths, Fig. 1,2) of the coccolithophorid phytoflagellates provides a particularly interesting model system for the study of biological mineralization, and the sequential formation of Golgi products.The coccoliths of Hymenomonas carterae consist of a scale-like base (Fig. 2 and 4, b) with a highly structured calcified (CaCO3) rim composed of two distinct elements which alternate about the base periphery (Fig. 1 and 3, A, B). Each element is enveloped by a sheath-like organic matrix (Fig. 3; Fig. 4, m).


Author(s):  
Douglas R. Keene ◽  
Gregory P. Lunstrum ◽  
Patricia Rousselle ◽  
Robert E. Burgeson

A mouse monoclonal antibody produced from collagenase digests of human amnion was used by LM and TEM to study the distribution and ultrastructural features of an antigen present in epithelial tissues and in cultured human keratinocytes, and by immunoaffinity chromatography to partially purify the antigen from keratinocyte cell culture media.By immunofluorescence microscopy, the antigen displays a tissue distribution similar to type VII collagen; positive staining of the epithelial basement membrane is seen in skin, oral mucosa, trachea, esophagus, cornea, amnion and lung. Images from rotary shadowed preparations isolated by affinity chromatography demonstrate a population of rod-like molecules 107 nm in length, having pronounced globular domains at each end. Polyacrylamide gel electrophoresis suggests that the size of this molecule is approximately 440kDa, and that it is composed of three nonidentical chains disulfide bonded together.


Author(s):  
Masako Osumi ◽  
Misuzu Nagano ◽  
Hiroko Kazama

We have found that microbodies appeared profusely together with a remarkable increase in catalase activity in normal alkane-grown cells of hydrocarbon-utilizing Candida yeasts, and that the microbodies multiplied by division in these cells. These features of Candida yeasts seem to provide a useful model system for studies on the biogenesis of the microbody. Subsequently, we have succeeded in isolation of Candida microbodies in an apparently native state, as judged biochemically and morphologically. The presence of DNA in the purified microbody fraction thus obtained was proved by the diphenylamine method. DNA molecule of about 15 urn in contour length was released from an isolated microbody. The physicochemical analyses of the microbody DNA revealed that its buoyant density differed from nuclear and mitochondrial DNAs. All these results lead us to the possibility that there is a novel type of DNA in microbodies.


Author(s):  
M.J. Witcomb ◽  
U. Dahmen ◽  
K.H. Westmacott

Cu-Cr age-hardening alloys are of interest as a model system for the investigation of fcc/bcc interface structures. Several past studies have investigated the morphology and interface structure of Cr precipitates in a Cu matrix (1-3) and good success has been achieved in understanding the crystallography and strain contrast of small needle-shaped precipitates. The present study investigates the effect of small amounts of phosphorous on the precipitation behavior of Cu-Cr alloys.The same Cu-0.3% Cr alloy as was used in earlier work was rolled to a thickness of 150 μm, solution treated in vacuum at 1050°C for 1h followed by quenching and annealing for various times at 820 and 863°C.Two laths and their corresponding diffraction patterns in an alloy aged 2h at 820°C are shown in correct relative orientation in Fig. 1. To within the limit of accuracy of the diffraction patterns the orientation relationship was that of Kurdjumov-Sachs (KS), i.e. parallel close-packed planes and directions.


Author(s):  
A. R. Crooker ◽  
M. C. Myers ◽  
T. L. Beard ◽  
E. S. Graham

Cell culture systems have become increasingly popular as a means of screening toxic agents and studying toxic mechanisms of drugs and other chemicals at the cellular and subcellular levels. These in vitro tests can be conducted rapidly in a broad range of relevant mammalian culture systems; a variety of biological and biochemical cytotoxicity endpoints can be examined. The following study utilized human keratinocytes to evaluate the relative cytotoxicities of nitrofurazone (NF) and silver sulfadiazine (SS), the active ingredients of FURACIN(R) Topical Cream and SILVADENE(R) Cream, respectively. These compounds are anti-infectives used in the treatment of burn patients. Cell ultrastructure and elemental composition were utilized as cytotoxicity endpoints.Normal Human Epidermal Keratinocytes (HK) were prepared from the EpiPackTM culture system (Clonetics Corporation, Boulder, CO). For scanning electron microscopy (SEM) and transmission electron microscopy (TEM), cells were seeded on sterile 35 mm Falcon plastic dishes; for elemental microanalysis, cells were plated on polished pyrolytic carbon discs (E. Fullam, Latham, NY) placed in the culture dishes.


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