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Human serum and epithelial spread in tissue culture

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Summary

The dependence of epithelial spread on human serum in culture has been studied. Using measured pieces of mouse ear skin epithelial outgrowth about floating explants (epiboly) and from adherent explants was studied. Where compared directly the two systems show similar results. Because of its ease of quantitation, the adherent explant culture was studied in greater detail. In this system in the presence of serum appreciable spread was found only after 48 h but spread continued for at least the next 4 days. In the absence of serum only minimal epithelial spread occurred. Adding serum to deficient media enhanced spreading and removing serum from media depressed spreading. The extent of spread appeared independent of mouse age for the first 4–10 weeks though 2-week-old mouse skin showed quantitatively greater spreading.

The activity in human serum responsible for epithelial spread acts under conditions of minimal DNA synthesis and is not reproduced by bovine serum albumin, fetuin, or serum that had been exposed to 100°C for 5 min. The activity is not dialyzable and it is resistant to the protease inhibitors DFP and PMSF. These studies suggest that a specific serum component(s) serves to support epithelial spread in vitro.

Zusammenfassung

Die Abhängigkeit von menschlichem Serum für den Auswuchs von Epithelzellen wurde in Gewebekultur an gemessenen Stücken von der Haut von Mausohren studiert. Ein direkter Vergleich von schwebenden und von anhaftenden Gewebestücken gab ähnliche Resultate. Das System mit anhaftenden Gewebestücken wurde ausführlicher studiert, da es einfache meßbare Daten gab. In diesem System wurde gefunden, daß Auswuchs von Epithelzellen in größerem Auswuchs nur nach 48 h erfolgt und zumindest für die nächsten 4 Tage anhielt. In Abwesenheit von Serum wurde geringerer Auswuchs beobachtet. Zugabe von Serum zur Gewebekultur erhöhte den Auswuchs, und Entfernung von Serum unterdrückte den Auswuchs der Epithelzellen. Das Ausmaß dieses Auswuchses war unabhängig vom Alter der Mäuse in den ersten 4–10 Wochen, obwohl Mäusehaut von 2 Wochen alten Tieren größeren Auswuchs zeigte. Die aktive Komponente in menschlichem Serum kann nicht ersetzt werden durch Rinderserum-Albumin und Fetuin, und wird zerstört durch Erhitzen für 5 min bei 100°C. Die aktive Serum-Komponente ist nicht dialysierbar, ist resistent für die Enzyminhibitoren DFP und PMSF und wirkt trotz geringer DNS-Synthese. Diese Studie deutet darauf hin, daß ein oder mehrere spezifische Serum-Komponenten den Auswuchs von Epithelzellen in Gewebekultur verursachen.

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Authors and Affiliations

  1. Department of Pathology, Yale School of Medicine, 06510, New Haven, CT, USA

    K. S. Stenn & I. Dvoretzky

  2. Department of Dermatology, Yale School of Medicine, 06510, New Haven, CT, USA

    K. S. Stenn & I. Dvoretzky

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  1. K. S. Stenn
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  2. I. Dvoretzky
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Stenn, K.S., Dvoretzky, I. Human serum and epithelial spread in tissue culture. Arch Dermatol Res 264, 3–15 (1979). https://doi.org/10.1007/BF00417274

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