Communication Between Normal Human Epidermal Cells in Vitro

Cavoto, Francis V.; Flaxman, B. Allen

doi:10.1111/1523-1747.ep12627491

Cited by 20 publications

(8 citation statements)

References 35 publications

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“…3) and long before the appearance of other transformed properties; e.g.,, growth independence from substrate attachment and from fibroblasts. Since growth and differentiation of epidermal cells have been postulated to be controlled by chemical gradients based upon direct (i.e., junctional) cell-to-cell communication (10,11), it is possible that through desmosomal complexes the mitogenic stimulation of the T antigen diffuses to neighboring noninfected cells.…”

Section: Characteristics Of Normal and Sv4o-infected Epidermalmentioning

confidence: 99%

Altered pattern of growth and differentiation in human keratinocytes infected by simian virus 40.

Steinberg¹,

Defendi²

1979

Proc. Natl. Acad. Sci. U.S.A.

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Human epidermal keratinocytes were infected by simian virus 40 in vitro. The structure of the developing keratinocyte colony reflects the spatial separation of cell division and keratinization in intact skin; thymidine-incorporating cells were primarily localized at the colony periphery whereas nondividing, histologically differentiated ce ls accumulated in the interior. Viral infection produced a dramatic increase in the size of the proliferative population as, simultaneously, differentiation was reduced in the colony interior. These changes were manifest when simian virus 40 T-antigen synthesis was detectable in only a small percentage of the cells; differentiation became increasingly density dependent as the percentage of T-antigen-positive cells rose over serial passage. The disruption of the normal pattern of growth/differentiation localization coincided with a loss of dependence on serum for growth, but preceded the appearance of other virus-induced properties associated with transformation; i.e., the ability to form colonies in soft agar and independence of growth from fibroblasts. Epidermis is a tissue in which differentiation is a process confined to a single cell type and defined geographically by the position of each cell in the tissue structure. These characteristics offer considerable advantages for the study of differentiation in comparison to other systems in which the developmental sequence is not directly related to tissue architecture or is dependent upon the influence of intervening effectors. In the past, research in this system has been impeded by the lack of adequate culture conditions for sustaining epidermal cell growth in vitro. Recently, however, a culture method has been developed that permits long-term serial cultivation of epidermal cells derived from human skin and that supports the growth of colonies probably arising from single cells (1). In this system the developmental process of epidermal architecture is maintained in terms of a geographical relationship between the replicating stem population and the terminally differentiating cells (1, 2).Using this system we have studied changes in the normal patterns of growth and differentiation resulting from infection of human epidermal cells by simian virus 40 (SV40). Our findings indicate that these processes are highly sensitive to viral infection and that modification of the normal pattern of growth and differentiation in the keratinocyte colonies are detectable prior to other parameters of cell transformation.METHODS AND MATERIALS Cell Culture and Virus Infection. Human epidermal keratinocyte cultures were prepared from tissue specimens as described by Rheinwald and Green (1). Briefly, freshly excised skin specimens of neonatal foreskin were washed in culture medium, minced with scissors, and trypsinized (0.25% trypsin, GIBCO) with rapid stirring for 45 min at room temperature. (70:30, vol/vol) and stained for T antigen prior to autoradiography. In some experiments, microscopic fields were projected onto paper screens wi...

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Section: Characteristics Of Normal and Sv4o-infected Epidermalmentioning

confidence: 99%

Altered pattern of growth and differentiation in human keratinocytes infected by simian virus 40.

Steinberg¹,

Defendi²

1979

Proc. Natl. Acad. Sci. U.S.A.

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“…The present study shows that electrotonic coupling is comparable for both normal and cancerous epidermal cells in vitro and that in both cases this is suggestive of low-resistance pathways . The distance over which electrotonic spread occurred is less than for some other epithelia, but may reflect specific tissue or environmental differences (Shiba, 1970(Shiba, , 1971 (Cavoto and Flaxman, 1972), but not between any cancers of epidermal origin .…”

Section: Discussionmentioning

confidence: 92%

“…Both the current pulse and transmembrane potential were displayed on the face of Tektronix 502A dual beam oscilloscope (Tektronix, Inc ., Beaverton, Ore.) and photographed with a Polaroid camera using 3000 ASA film . The methods of measuring electrotonic coupling were standard and are described in detail elsewhere (Cavoto and Flaxman, 1972) .…”

Section: Methodsmentioning

confidence: 99%

“…Studies of cancer cells, where coupling might be expected to be absent or diminished when compared with normal cells, have given conflicting results . Both normal and transformed fibroblasts have been found to be coupled in vitro (Potter et al ., 1966 ;Furshpan and Potter, 1968 ;Borek et al ., 1969) and it might be concluded that this aspect of cell function per se is not related to those aspects of a fibroblast's behavior that give it a "cancerous" Electrotonic coupling has been demonstrated between normal cells from a variety of tissues (see reviews by Loewenstein, 1966 ;Furshpan and Potter, 1968 ;Sheridan, 1970 ;Cavoto andFlaxman, 1972, Van Heukelom et al ., 1972 b), but the significance of this type of coupling is not yet clear . The passage of charged molecules from cell to cell implies a role in regulating some aspects of behavior, e .g.…”

Section: Introductionmentioning

confidence: 99%

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Low-Resistance Junctions in Epithelial Outgrowths From Normal and Cancerous Epidermis in Vitro

Flaxman

Cavoto

1973

The Journal of Cell Biology

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“…With contact, electronic coupling occurs (Furshpan & Potter, 1968 ;Gilula et al, 1972 ;Johnson & Sheridan, 1971;Cavoto & Flaxman, 1972). Although dividing cells (in vitro) appear to lose intimate contact with surrounding interphase cells when examined by light microscopy, O'Lague et al (1970) have shown in fibroblasts in vitro, and Cavoto & Flaxman (1973)have shown in epithelial cells in vitro that electronic coupling persists.…”

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confidence: 99%

Biology in vitro of corneal epithelium and endothelium

Yanoff

1976

Doc Ophthalmol

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Four main areas were explored: 1) the proper medium for culturing corneal tissue; 2) the effect of serum on tissue growth in vitro; 3) the interrelationships in vitro between corneal epithelium and endothelium, and 4) the biology of cultures of whole corneas (organ cultures). Modified Eagle's minimal essential medium (MEM) proved to be an excellent culture fluid. Corneal tissue could be grown in MEM without serum or clot, thus providing a defined culture medium. The biology in vitro of outgrowths of multilayered corneal epithelium and monolayered corneal endothelium are discussed. Contact inhibition between epithelium and endothelium is demonstrated in whole corneal (organ) cultures.

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Communication Between Normal Human Epidermal Cells in Vitro

Cited by 20 publications

References 35 publications

Altered pattern of growth and differentiation in human keratinocytes infected by simian virus 40.

Altered pattern of growth and differentiation in human keratinocytes infected by simian virus 40.

Low-Resistance Junctions in Epithelial Outgrowths From Normal and Cancerous Epidermis in Vitro

Biology in vitro of corneal epithelium and endothelium

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