Epidermal growth factor and the multiplication of cultured human epidermal keratinocytes

Rheinwald, James G.; Green, Howard

doi:10.1038/265421a0

Cited by 1,008 publications

(461 citation statements)

References 24 publications

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“…Cell Culture-Punch biopsies obtained from the probands were used to establish primary cultures on lethally irradiated murine 3T3 fibroblasts as described earlier (37)(38)(39). Secondary cultures were grown in high calcium keratinocyte medium, 10% fetal calf serum until confluency.…”

Section: Methodsmentioning

confidence: 99%

Consequences of Seven Novel Mutations on the Expression and Structure of Keratinocyte Transglutaminase

Huber¹,

Yee²,

Burri³

et al. 1997

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

Consequences of Seven Novel Mutations on the Expression and Structure of Keratinocyte Transglutaminase

Huber¹,

Yee²,

Burri³

et al. 1997

Journal of Biological Chemistry

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“…They were grown according to the procedure of Rheinwald and Green (20,21). When vitamin A is present in the cell culture medium, terminal differentiation is largely inhibited, and the cells resemble basal epidermal ceils (22).…”

Section: Extraction Of Keratins and Poly(a)+ Rnas From Culturedmentioning

confidence: 99%

Expression of unusually large keratins during terminal differentiation: balance of type I and type II keratins is not disrupted.

Kim¹,

Marchuk²,

Fuchs³

1984

The Journal of Cell Biology

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When a basal epidermal cell undergoes a commitment to terminally differentiate, it ceases to divide and begins to migrate outward towards the surface of the skin. Dramatic changes in its cytoskeletal architecture take place, accompanied by numerous changes in the expression of keratins, a family of related polypeptides that form 8-nm filaments in these cells. We show here that a shift to the synthesis of unusually large keratins occurs that does not seem to disrupt the ratio of two distinct subfamilies of keratins. Preliminary studies indicate that this differentiation-specific shift may be at the level of transcriptional rather than post-trancriptional regulation. The striking similarities between these large keratins and the type I and type II keratins of basal epidermal cells suggests the important role that both classes of large keratin sequences must play in the assembly of the intermediate filaments within the differentiating keratinocyte.Several laboratories have reported differences between the keratins of stratum corneum and those of the living layers of epidermis (1--4). During the course of terminal differentiation, changes occur in the expression of the keratin polypeptides that comprise the 8-nm tonofilaments (5-10). For the human, keratins of size 46, 48.5, 50, 52, 56, and 58 kd are synthesized by the basal epidermal cells (Fig. 1, lane 1), whereas additional keratins of size 67, 65.5, and 56.5 kd are produced by the terminally differentiating epidermis (lane 3). As the cells pass through the granular layer to the stratum corneum, a slight reduction in the size of the keratins takes place. As judged by in vitro translation of mRNAs isolated from basal (lane 2) and differentiating (lane 4) keratinocytes, the changes in keratin pattern that occur early in the course of terminal differentiation are clearly at the level of mRNA biosynthesis (5).Recently, it was demonstrated that the keratins produced by basal epidermal cells can be divided into two distinct groups based on the ability of their mRNAs to cross-hybridize with two different cloned keratin cDNAs (11). Other epithelia express different subsets of keratins, but most if not all of these seem to be similar to one or the other of the two epidermal keratin subfamilies (12). The small (40-52 kd) and relatively acidic keratins have been named the type I class, and the large (53-58 kd) and more basic keratins have been named the type II class (13,14). At least one member of each of the two keratin types seem to be expressed in all cells at all times, suggesting their combined importance in filament assembly (14). Sequence analyses (13,(15)(16)(17)(18) have shown that while the two types of keratins share only low (<30%) homology, their predicted secondary structures are strikingly similar and are compatible with their playing an essential role in forming the coiled-coil backbone of the protofilament of the 8-nm keratin filament (13,18).The keratins typical of terminally differentiating keratinocytes seem to be unusually large and are not fo...

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“…In contrast, cells incubated with 10 ng of EGF/mL or 10% FBS showed intermediate migration when compared to those incubated with both ( Figures 2B-D). The effect of EGF and FBS on BALB/MK cell growth differed: cells incubated with EGF were more dispersed (indicating motogenic activity) whereas those incubated with FBS formed a dense layer (indicating greater mitogenicity and less motogenicity), as previously reported for primary cultures of keratinocytes (Rheinwald and Green, 1977;Barrandon and Green, 1987).…”

Section: Effects Of Egf On Balb/mk Cells Modified With Lesnmentioning

confidence: 57%

Autonomous growth of BALB/MK keratinocytes transfected with a retroviral vector carrying the human epidermal growth factor gene

et al. 2008

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Epidermal growth factor (EGF), which promotes epidermal regeneration and wound closure, is important for the proliferation and differentiation of epidermal and epithelial tissues in animals. Exogenous EGF is a promising therapeutic agent for wound healing, but its general use is restricted by the limited availability of this protein. In this work, we show that the transfection of mouse BALB/MK keratinocytes, which are totally dependent on EGF for growth and migration, with mature cDNA for human EGF via a retroviral vector abolished the cells requirement for exogenous EGF. The transformed cells had normal morphology and a growth rate that varied according to the source of the retroviral vector used. Keratinocyte transfection with EGF cDNA provides a time-and cost-efficient means of culturing keratinocytes and yields cells that may be useful for skin grafting.

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Epidermal growth factor and the multiplication of cultured human epidermal keratinocytes

Cited by 1,008 publications

References 24 publications

Consequences of Seven Novel Mutations on the Expression and Structure of Keratinocyte Transglutaminase

Consequences of Seven Novel Mutations on the Expression and Structure of Keratinocyte Transglutaminase

Expression of unusually large keratins during terminal differentiation: balance of type I and type II keratins is not disrupted.

Autonomous growth of BALB/MK keratinocytes transfected with a retroviral vector carrying the human epidermal growth factor gene

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