Study of epidermal differentiation in human keratinocytes cultured in autocrine conditions. 2010

Frédéric Minner, and Françoise Herphelin, and Yves Poumay
Cell and Tissue Laboratory, URPHYM, University of Namur (FUNDP), Namur, Belgium.

This chapter deals with protocols to set up human keratinocyte cultures in serum-free conditions and lead them to autocrine autonomously growing conditions. These conditions have proven adequate for studies of epidermal differentiation by measurements of the expression of typical early and late differentiation markers. The chapter also deals with the use of quantitative RT-PCR in order to determine the epidermal marker gene expression levels by comparison with adequate housekeeping genes.

UI MeSH Term Description Entries
D002454 Cell Differentiation Progressive restriction of the developmental potential and increasing specialization of function that leads to the formation of specialized cells, tissues, and organs. Differentiation, Cell,Cell Differentiations,Differentiations, Cell
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D004817 Epidermis The external, nonvascular layer of the skin. It is made up, from within outward, of five layers of EPITHELIUM: (1) basal layer (stratum basale epidermidis); (2) spinous layer (stratum spinosum epidermidis); (3) granular layer (stratum granulosum epidermidis); (4) clear layer (stratum lucidum epidermidis); and (5) horny layer (stratum corneum epidermidis).
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000078404 Epidermal Cells Cells from the outermost, non-vascular layer (EPIDERMIS) of the skin. Epidermal Cell,Epidermic Cells,Cell, Epidermal,Cell, Epidermic,Cells, Epidermic,Epidermic Cell
D015603 Keratinocytes Epidermal cells which synthesize keratin and undergo characteristic changes as they move upward from the basal layers of the epidermis to the cornified (horny) layer of the skin. Successive stages of differentiation of the keratinocytes forming the epidermal layers are basal cell, spinous or prickle cell, and the granular cell. Keratinocyte
D015870 Gene Expression The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION. Expression, Gene,Expressions, Gene,Gene Expressions
D016895 Culture Media, Serum-Free CULTURE MEDIA free of serum proteins but including the minimal essential substances required for cell growth. This type of medium avoids the presence of extraneous substances that may affect cell proliferation or unwanted activation of cells. Protein-Free Media,Serum-Free Media,Low-Serum Media,Culture Media, Serum Free,Low Serum Media,Media, Low-Serum,Media, Protein-Free,Media, Serum-Free,Media, Serum-Free Culture,Protein Free Media,Serum Free Media,Serum-Free Culture Media
D019898 Autocrine Communication Mode of communication wherein a bound hormone affects the function of the cell type that produced the hormone. Autocrine Signaling,Autocrine Communications,Autocrine Signalings,Communication, Autocrine,Communications, Autocrine,Signaling, Autocrine,Signalings, Autocrine
D020133 Reverse Transcriptase Polymerase Chain Reaction A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols. Polymerase Chain Reaction, Reverse Transcriptase,Reverse Transcriptase PCR,PCR, Reverse Transcriptase,Transcriptase PCR, Reverse

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