Cell
Volume 22, Issue 2, Part 2, November 1980, Pages 629-632
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Article
Defective terminal differentiation in culture as a consistent and selectable character of malignant human keratinocytes

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Abstract

Culture conditions can be manipulated to vary the rate at which keratinocytes (stratified squamous epithelial cells) become committed to differentiate terminally. When deprived of anchorage in semi-solid medium, normal human keratinocytes irreversibly lose the ability to reinitiate growth in surface culture with a t12 of 3 hr and then proceed to form cornified envelopes. We examined six established lines from human squamous cell carcinomas (SCCs) for defects in this function. One SCC line which grew progressively in semi-solid medium could not be induced to form cornified envelopes. The other five lines, which grew abortively, at best, in semi-solid medium, formed envelopes under this condition but at subnormal rates. During anchorage deprivation the SCC lines became committed to differentiate much more slowly than normal, with t12's of 24–144 hr. SCC cells therefore possess at least a partial defect in the triggering of terminal differentiation. In vivo, such an alteration may permit malignant behavior by evading an important tissue-specific mechanism for limiting growth. In culture, the phenotype of increased survival in semi-solid medium may be used to detect and select malignantly transformed keratinocytes.

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    IHOK were provided by the Department of Oral Pathology and Oral Cancer Research Institute at Yonsei University College of Dentistry (Seoul, Korea) [36]. HSC-2 (JCRB0622, Japanese Collection of Research Bioresources, Japan) and SCC-15 (CRL-1623, American Type Culture Collection, USA) cells were purchased and used as representative epithelial-like and spindle-like OSCC keratinocytes, respectively [37,38]. IHOK and HSC-2 were incubated in a DMEM/F-12 3:1 mixture media (Welgene, Daegu, Korea), and SCC-15 was incubated in a DMEM/F-12 1:1 mixture media (Welgene).

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