Use of Immortalized Human Keratinocytes for the Study of Squamous Differentiation and Mutagenesis

Inactivation of p53 in Human Keratinocytes Leads to Squamous Differentiation and Shedding via Replication Stress and Mitotic Slippage

Cell Reports ◽

10.1016/j.celrep.2014.10.012 ◽

2014 ◽

Vol 9 (4) ◽

pp. 1349-1360 ◽

Cited By ~ 31

Author(s):

Ana Freije ◽

Rut Molinuevo ◽

Laura Ceballos ◽

Marta Cagigas ◽

Pilar Alonso-Lecue ◽

...

Keyword(s):

Replication Stress ◽

Human Keratinocytes ◽

Squamous Differentiation ◽

Mitotic Slippage

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Kalinin: A new epithelial basement membrane adhesion molecule

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100085198 ◽

1991 ◽

Vol 49 ◽

pp. 178-179

Author(s):

Douglas R. Keene ◽

Gregory P. Lunstrum ◽

Patricia Rousselle ◽

Robert E. Burgeson

Keyword(s):

Basement Membrane ◽

Culture Media ◽

Polyacrylamide Gel Electrophoresis ◽

Human Keratinocytes ◽

Positive Staining ◽

Human Amnion ◽

Epithelial Basement Membrane ◽

Type Vii Collagen ◽

Keratinocyte Cell ◽

Epithelial Basement

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.

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Alterations of ultrastructure and elemental composition in cultured human epidermal keratinocytes after treatment with nitrofurazone and silver sulfadiazine

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100127803 ◽

1987 ◽

Vol 45 ◽

pp. 676-677

Author(s):

A. R. Crooker ◽

M. C. Myers ◽

T. L. Beard ◽

E. S. Graham

Keyword(s):

Electron Microscopy ◽

Elemental Composition ◽

Pyrolytic Carbon ◽

Human Keratinocytes ◽

Silver Sulfadiazine ◽

Epidermal Keratinocytes ◽

Human Epidermal Keratinocytes ◽

Culture Systems ◽

Cytotoxicity Endpoints

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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