Effects of retinoic acid and other retinoids on the gowth and differentiation of 3t3 supported human keratinocytes

Kubilus, Joseph; Rand, Rhonda; Baden, Howard P.

doi:10.1007/bf02618445

Cited by 31 publications

(13 citation statements)

References 37 publications

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“…The decrease in the amount of tonofilaments and the formation of wide and loosely packed bundles of tonofilaments seen in the retinoid-treated cultures of the present study have also been reported in retinoid-treated skin explants [l]. Retinoids induce a decrease in the ratio of prekeratin and keratin to the total solubilizable proteins in treated cultures [21]. Retinoids also inhibit the synthesis of high-molecular-weight keratins, promote the synthesis of low-molecular-weight keratins in culture [lo], and inhibit the cross-linking of keratins [34].…”

Section: Discussionsupporting

confidence: 87%

Retinoids alter the direction of differentiation in primary cultures of cutaneous keratinocytes

1985

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The effects of vitamin A on the morphological expression of differentiation were studied in cell cultures of cutaneous keratinocytes from the newborn rat. The cells were first cultivated in a medium containing 0.11 mM calcium until a confluent monolayer had been formed. Stratification and terminal differentiation were then triggered by raising the calcium concentration of the medium to 1.96 mM ('normal' culture). The rise in the concentration of calcium was coupled with the addition of retinol (RL) of retinoic acid (RAC) to the medium to produce an excess of vitamin A (high-retinoid culture). Delipidized serum was used to produce a deficiency of vitamin A (low-retinoid culture). The tissue organization and the ultrastructure of the keratinocytes in the stratified culture were the same as those seen in conventional cultures and skin explants. These stratified cultures expressed the morphological features of the epidermis of intact skin. The addition of RL or RAC to the medium enhanced features characteristic of the secretory epithelium, such as the formation of an extensive endoplasmic reticulum, an enlargement of the Golgi zone, and an increase in the number of vacuoles. At the same time, the addition of retinoids diminished features characteristic of the terminal differentiation of the stratified squamous epithelium, such as stratification and keratinization. Deficiency of vitamin A in the medium resulted in a culture with many differentiated layers. The differentiated cells of the low-retinoid cultures contained densely packed tonofilaments and synthesized products that reacted with the monoclonal antibody AE2 that is specific for keratin peptides which are markers of epidermal differentiation. In the cell culture system that is presented here, an excess of retinoids redirected epithelial differentiation from a stratifying and keratinizing epithelium towards a secretory epithelium. This system is a useful tool for elucidating the mechanisms responsible for the effect of vitamin A on the differentiation of epithelial cells.

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Section: Discussionsupporting

confidence: 87%

Retinoids alter the direction of differentiation in primary cultures of cutaneous keratinocytes

1985

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“…Conversely, TJ assembly could be dependent on cytoskeletal network and desmosomal organization, which may be altered by RA as previously shown (Kubilus et al, 1981; Kopan et al, 1987; Debal et al, 1997; Wanner et al, 1999; Hatakeyama et al, 2004). Supporting this alternative, examination of the cultured epithelia treated with 10 µM RA showed that epithelial barrier disruption was associated to increased intercellular spaces and a reduced desmosome number (supplementary material Fig.…”

Section: Discussionmentioning

confidence: 60%

Expression of claudins -2 and -4 and cingulin is coordinated with the start of stratification and differentiation in corneal epithelial cells: retinoic acid reversibly disrupts epithelial barrier

et al. 2012

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SummaryAlthough tight junctions (TJ) have been extensively studied in simple epithelial cells, it is still unknown whether their organization is coupled to cell differentiation in stratified epithelia. We studied the expression of TJ in RCE1(5T5) cells, an in vitro model which mimics the sequential steps of rabbit corneal epithelial differentiation. RCE1(5T5) cells expressed TJ components which were assembled once cells constituted differentiated epithelia, as suggested by the increase of transepithelial electrical resistance (TER) which followed a similar kinetic to the expression of the early differentiation marker Pax-6. TJ were functional as indicated by the establishment of an epithelial barrier nonpermeable to ruthenium red or a biotin tracer. In immunostaining experiments, TJ were located at the superficial cells from the suprabasal layers; Western blot and RT-PCR suggested that TJ were composed of claudins (cldn) -1, -2, -4, cingulin (cgn), occludin (ocln) and ZO-1. Semi-quantitative RT-PCR and TER measurements showed that TJ became organized when cells began to form a 3–5 layers stratified epithelium; TER increased once cells reached confluence, with a time course comparable to the raise in the expression of cgn, cldn-2 and -4. Nevertheless, cldn-1, -2, ZO-1 and ocln were present in the cells from the beginning of cultivation, suggesting that TER increases mainly depend on TJ assembly. While EGF increased epithelial barrier strength, retinoic acid disrupted it, increasing paracellular flux about 2-fold; this effect was concentration dependent and completely reversible. Our results suggest that TJ assembly is tightly linked to the expression of corneal epithelial terminal phenotype.

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“…Vitamin A-sufficient cells dissolved the collagen substrate substantially faster than did deficient (A-) cells, and the collagenolytic activity of the deficient cells was partially restored by addition of 10-7M retinoic acid to the culture medium. A large body of evidence suggests that vitamin A modulates epithelial cell differentiation in vivo as well as in vitro (Wolbach and Howe, 1925;Fuchs and Green, 1981;Kubilus et al, 1981;Strickland and Mahdavi, 1978;Chopra and Flaxman, 1975). Fuchs and Green (1981) and Kim et al (1984) have previously shown that removal of vitamin A from the medium of cultured human skin keratinocytes induces expression of a Mr = 67 K keratin characteristic of the terminally differentiated state, and that this effect is reversed by addition of retinoids.…”

Section: Discussionmentioning

confidence: 99%

“…Vitamin A and its structural analogues (retinoids) have multiple biological effects in addition to their well-documented role in the visual process, and a body of evidence suggests that vitamin A is required in vivo for normal growth and differentiation of diverse types of cells, including several mammalian epithelia (Kubilus et al, 1981;Dowling and Wald, 1960;Wolbach and Howe, 1925;Lotan, 1980;Wolf, 1980). The mechanism of action of vitamin A on epithelial cells is still poorly understood, but it is of note that retinoids have been proven useful in the treatment of a variety of skin disorders in humans (Fulton, 1975;Fulton et al, 1968;Kligman et al, 1969;GUnther, 1973), and of experimentally induced tumors in animals (Bollag, 1974;Sporn et al, 1977).…”

Section: Introductionmentioning

confidence: 99%

Basic Biological Sciences

Wells

Birkedal‐Hansen

1985

J Dent Res

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A large body of evidence suggests that retinoids modulate the phenotypic expression of epithelial cells of skin and mucous membranes. The purpose of this study was to investigate the effect of retinoic acid on keratinocyte-mediated collagen breakdown. Keratinocytes derived from the ventral surfaces of the tongues of 4-6 week-old male Wistar rats were established in culture under conditions which are restrictive to growth of fibroblasts, and they were eventually cloned by limiting dilution. The cells were seeded (100,000 cells/cm2) in dishes coated with 3H-labeled, reconstituted type I collagen fibrils and incubated in serum-free medium over a 3-5 day period. Dissolution of the collagen fibrils was monitored by the release of radioactivity to the culture medium. Unstimulated cells metabolized the collagen rather slowly, but addition of retinoic acid in concentrations from 10(-6)M to 10(-8)M resulted in marked acceleration of the degradative process, with complete solubilization of the collagen fibrils in four or five days. The effect of 10(-6)M retinoic acid was of the same order of magnitude as that obtained by addition of a proteolytic activating system either in the form of plasmin or of plasminogen, which is converted to catalytic plasmin by endogenous activators. The effects of retinoic acid and plasminogen/plasmin, however, were not additive. Keratinocytes rendered vitamin A-deficient by cultivation in sera from deficient rats were clearly less effective in degrading the collagen substrate than were "sufficient" cells. Addition of retinoic acid (10(-7)M) enhanced collagen breakdown in both sets of cultures and partially restored the collagenolytic activity of the deficient cells.

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Effects of retinoic acid and other retinoids on the gowth and differentiation of 3t3 supported human keratinocytes

Cited by 31 publications

References 37 publications

Retinoids alter the direction of differentiation in primary cultures of cutaneous keratinocytes

Retinoids alter the direction of differentiation in primary cultures of cutaneous keratinocytes

Expression of claudins -2 and -4 and cingulin is coordinated with the start of stratification and differentiation in corneal epithelial cells: retinoic acid reversibly disrupts epithelial barrier

Basic Biological Sciences

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