Adult human keratinocyte cultures express 40, 52, 58 and 67 KD keratins
Keratins are complex fibrous proteins characteristic of epithelial cells. Although several different classes of keratins are known to occur in the epidermis, the expression of all keratins has not been observed in vitro. The authors have developed a procedure that allows us to culture and passage up to ten times, adult human kerationcytes, in the absence of mesenchymal substrates. EM examination of stratifying cultures showed the presence of numerous tonofilaments, desmosomes and keratohyaline granules. The expression of different classes of keratins was examined by immunofluorescence, radiolabeling, SDS-PAGE and Western blot, using mouse monoclonal antibodies. Analysis of water-insoluble proteins showed the presence of keratins of M.W. 40 kd, 50-52 kd, 56-57 kd and 65-67 kd. The expression of 40kd keratin is known to be associated with basal cells. In their culture system basal cells secrete a well-defined basement membrane on which they rest. These cells may be responsible for the 40kd keratin. The expression of 65-67kd keratins has not previously been observed in vitro. These keratins are considered to be markers for terminal differentiation of epidermal cells. These proteins are presumed to be synthesized in their cultures by sloughing layers of rough, granular cells.
- Research Organization:
- Univ. of California, San Francisco
- OSTI ID:
- 5462745
- Report Number(s):
- CONF-8604222-; TRN: 86-028377
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
EPIDERMIS
CELL DIFFERENTIATION
KERATIN
BIOSYNTHESIS
CELL CULTURES
CELL MEMBRANES
EPITHELIUM
FLUORESCENCE
IN VITRO
MAN
MICE
MONOCLONAL ANTIBODIES
RADIOISOTOPES
TRACER TECHNIQUES
ANIMAL TISSUES
ANIMALS
ANTIBODIES
BODY
CELL CONSTITUENTS
ISOTOPE APPLICATIONS
ISOTOPES
LUMINESCENCE
MAMMALS
MEMBRANES
ORGANIC COMPOUNDS
ORGANS
PRIMATES
PROTEINS
RODENTS
SCLEROPROTEINS
SKIN
SYNTHESIS
TISSUES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques