title: Purification of porcine hair keratin subunits and their immobilization for use as cell culture substrates.
abstract: We purified both the type I subunit and type II subunit of porcine hair keratin and compared their ability to form a uniform film of reconstituted keratin on a culture plate, and their effect on a model of neural cells. We observed the surface of the keratin-immobilized plate using a scanning electron microscope (SEM) and measured water contact angles to characterize the surface. We cultured PC12 cells on plates on which crude keratin, the type I subunit, or the type II subunit were immobilized. The water contact angles were slightly different from each other. The cells proliferated well on all three keratin-immobilized plates. The type II subunit showed a tendency to inhibit the differentiation of PC12 cells significantly as an extension of the cell shapes and neurite outgrowth in comparison with the crude extract and the type I subunit. The type I subunit and the type II subunit showed slight differences in cell differentiation, but not in cell proliferation.
authors.affiliation: Department of Biological Science, Graduate School of Science, Osaka Prefecture University.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Ozaki
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Takagi
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Saito
authors.forename: Hideki
authors.initials: H
authors.lastname: Mori
authors.forename: Masayuki
authors.initials: M
authors.lastname: Hara
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D006197:Hair / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D007633:Keratins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D053530:Keratins, Type I / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D053531:Keratins, Type II / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D008969:Molecular Sequence Data; D016716:PC12 Cells; D055550:Protein Stability; D051381:Rats; D013499:Surface Properties; D013552:Swine
chemical_list: D055767:Immobilized Proteins; D053530:Keratins, Type I; D053531:Keratins, Type II; D007633:Keratins