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    • 4. 发明申请
    • IMMORTALISED CELL LINES DERIVED FROM NORMAL HUMAN SKIN TISSUES
    • 来自正常人类皮肤组织的免疫细胞系
    • WO9954435A2
    • 1999-10-28
    • PCT/EP9902347
    • 1999-04-07
    • SOC D PRODUITS NESTLEBAUR MARKUS
    • BAUR MARKUS
    • C12N15/09A61L27/00C12N1/00C12N5/071C12N5/10C12N
    • C12N5/0629C12N2500/14C12N2500/38C12N2501/11C12N2503/06C12N2510/04
    • The invention concerns a cell line of human keratinocytes immortalised by at least a functional tumorigenic gene derived from a retrovirus characterised in that it is (1) non-tumorigenic, (2) preserves aptitude for differentiating and expressing proteins and enzymes which are expressed by normal differentiated keratinocytes even after a high rate of passages in tissue culture, and (3) forms a stratified and polarised epithelium comprising an ortho-keratotic stratum corneum, when cultivated in organotypical culture in a medium without serum and without feeder cell layer. The invention also concerns an improved method for immortalising human skin cells to obtain immortalised keratinocytes, characterised in that it comprises a step of infecting the keratinocytes using functional tumorigenic genes of at least two different retoviruses. The invention further concerns the use of said keratinocytes for immunologic, pharmacological, photo-toxicological and chemiotoxicological skin reactions for expressing heterologous genes and for constructing an artificial skin.
    • 本发明涉及由至少一种源自逆转录病毒的功能性致瘤基因永生化的人角质形成细胞的细胞系,其特征在于它是(1)非致瘤性的,(2)保持分化和表达蛋白质的能力和由正常 即使在组织培养中传代率高时分化的角质形成细胞,以及(3)当在没有血清且没有饲养细胞层的培养基中在器官培养中培养时,形成包含角膜角化正常角质层的分层和极化上皮。 本发明还涉及使人皮肤细胞永生化以获得永生化角质形成细胞的改进方法,其特征在于它包括使用至少两种不同的逆转录病毒的功能致瘤基因感染角质形成细胞的步骤。 本发明还涉及所述角质形成细胞用于表达异源基因和构建人造皮肤的免疫学,药理学,光毒理学和化学毒性皮肤反应的用途。