In preferred embodiments, the activity of variant or mutant GKLF is evaluated by the methods described above. In this case, the GKLF encoding polynucleotide sequences are typically employed in an aqueous solution containing salts e. In some embodiments, the vector is an episomal vector such as a plasmid. The Klf4 cDNA is also cloned into an expression plasmid containing promoter sequences from the involucrin gene. USA, 90 21 : p. Cultures that had previously received base media with additional constituents supplemented with 0.
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In some embodiments, the methods comprise providing keratinocytes and a culture media comprising ascorbic acid and linoleic acid; and culturing the keratinocytes under conditions such that a skin equivalent having improved barrier function is formed.
Geiman et al.
The equation for calculating the T m of nucleic acids is well known in the art. Thus they were unsuitable for commercial application. For example, a group of amino acids having aliphatic side chains is glycine, alanine, valine, leucine, and isoleucine; a group of amino acids having aliphatic-hydroxyl side chains is serine and threonine; a group of amino acids having amide-containing side chains is asparagine and glutamine; a group of amino acids having aromatic side chains is phenylalanine, tyrosine, and tryptophan; a group of amino acids having basic side chains is lysine, arginine, and histidine; and a group of amino acids having sulfur-containing side chains is cysteine and methionine.
These methods may be adapted to study the delivery of drugs via transdermal patches or other delivery modes. In some embodiments, the skin equivalents are mounted on top of modified Franz diffusion cells.
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Stable NIKS cell lines that express Klf4 from the involucrin promoter or the doxycycline-inducible system are examined in organotypic culture to confirm that Klf4 is expressed under these culture conditions. Two days post-plating, the organotypic cultures were supplied with fresh medium to maintain growth.
For example, growing the cultures at lowered relative humidity improves the barrier function of skin equivalent cultures Mak et al.
With medium stringency conditions, nucleic acid base pairing will occur between nucleic acids with an intermediate frequency of complementary base sequences e.
Impedance meter readings were used to measure the resulting barrier function of cultures. These gene expression changes indicate that one or more regulatory transcription factors are responsible for altering the gene expression profile of differentiating keratinocytes to facilitate development of the epidermal barrier. A sample suspected of containing a human chromosome or sequences associated with a human chromosome may comprise a cell, chromosomes isolated from a cell e.
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|In still further preferred embodiments, linoleic acid is provided at a concentration of from about 5 to 80 micromolar.
Zhang et al. Baden, In Vitro cell. To determine whether preparation of organotypic skin cultures using the culture conditions shown in Table 2 has a synergistic effect on epidermal lipid composition, the lipid profiles of supplemented cultures are compared to lipids isolated from unsupplemented control cultures and to lipids extracted from normal human skin.
Complex developmental programs can be initiated by the action of one or a small number of key regulatory transcription factors, sometimes called master regulators or selector genes. Gene Therapy In still further embodiments, the skin equivalent is engineered to provide a therapeutic agent to a subject.
Primary Class: .Academic Press, Inc.: San Diego (). Stratech stems from UW-Madison research lead to Stratatech, the Madison- based skin regeneration company she founded in She visited the regional burn center at University of Wisconsin School of Medicine and.
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Madison College Biotechnology Stratech Corporation, Trac Microbiology, University of Wisconsin Biotech Center, University of Madison WI
It is believed that transepidermal water flux may serve as a regulatory signal for epidermal lipid synthesis and repair following disruption of the epidermal barrier Grubauer et al.
Cosmetic companies are interested in finding formulations that improve the barrier function. In other embodiments, DNA sequences derived from the SV40 splice, and polyadenylation sites may be used to provide the required non-transcribed genetic elements. The reference sequence may be a subset of a larger sequence, for example, as a segment of the full-length sequences of the compositions claimed in the present invention e. Cultures of differentiated keratinocytes are deficient in several ceramides that are major constituents of normal skin Ponec et al.
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|In contrast to the defects caused in other mutant mouse lines, the absence of Klf4 does not result in gross alterations of the epidermal ultrastructure or in lipid profiles.
In normal human epidermis, as the basal cells divide they give rise to daughter cells that migrate upwards into the differentiating layers of the tissue. Tim Cullen wins national award for increasing teacher diversity. Development of the epidermal permeability barrier requires the coordinated synthesis and metabolism of keratinocyte-specific protein and lipid products that are assembled into the outermost skin layer, the stratum corneum.
These seeded skin equivalents can then be used to screen compounds or other treatment strategies e. This example describes the effect of prelifting for the entire life of the dermal equivalent. The presence of these structures was especially evident in areas where the parental cells or the NIKS cells had attached directly to the porous support.