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WO2000050892A1 - Modele in vivo et in vitro pour photovieillissement cutane et destruction oxydative - Google Patents

Modele in vivo et in vitro pour photovieillissement cutane et destruction oxydative Download PDF

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WO2000050892A1
WO2000050892A1 PCT/US2000/004703 US0004703W WO0050892A1 WO 2000050892 A1 WO2000050892 A1 WO 2000050892A1 US 0004703 W US0004703 W US 0004703W WO 0050892 A1 WO0050892 A1 WO 0050892A1
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mouse
fibroblast culture
elastin promoter
human elastin
transgenic
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Eric F. Bernstein
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Priority to EP00911941A priority patent/EP1163514A4/fr
Priority to JP2000601437A priority patent/JP2002542764A/ja
Priority to AU33754/00A priority patent/AU761728B2/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/027—New or modified breeds of vertebrates
    • A01K67/0275—Genetically modified vertebrates, e.g. transgenic
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/027—New or modified breeds of vertebrates
    • A01K67/0275—Genetically modified vertebrates, e.g. transgenic
    • A01K67/0278—Knock-in vertebrates, e.g. humanised vertebrates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
    • A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2207/00—Modified animals
    • A01K2207/15—Humanized animals
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2227/00—Animals characterised by species
    • A01K2227/10—Mammal
    • A01K2227/105—Murine
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2267/00—Animals characterised by purpose
    • A01K2267/03—Animal model, e.g. for test or diseases
    • A01K2267/0393—Animal model comprising a reporter system for screening tests

Definitions

  • Elevated elastin mRNA levels in sun- damaged skin result from enhanced elastin promoter activity, as shown by transient transfections of fibroblasts with a DNA construct composed of the human elastin promoter linked to the chloramphenicol acetyltransferase (CAT) reporter gene (Bernstein et al . , J. Invest . Dermatol . , 1994, 103:182-186).
  • CAT chloramphenicol acetyltransferase
  • a transgenic mouse line expressing the 5.2 kb human elastin promoter linked to a chloramphenicol acetyltransferase reporter gene (CAT) has been developed which models cutaneous photoaging (Bernstein et al . , J. Invest . Dermatol . , 1995, 105, 269-273). Although phenotypically normal, the cells in these mice possess the human elastin promoter/CAT construct, allowing elastin promoter activity to be measured in response to stimuli such as ultraviolet radiation (UV) .
  • UV ultraviolet radiation
  • mice which have not yet developed hair, and fibroblast cultures derived from their skin, have been demonstrated to provide a rapid and sensitive means of identifying compounds capable of inhibiting cutaneous photodamage (Bernstein et al . , J. Invest . Derma tol . , 1995, 105, 269-273; Bernstein et al . , Photochem . Photobiol . , 1996, 64:369-74; Bernstein et al . , J " . Am . Acad . Dermatol . , 1997, 37:725-729).
  • UV from the sun also damages skin by the generation of reactive oxygen species (Miyachi, Y. J., Dermatol . Sci . , 1995, 9:79-86) .
  • Reactive oxygen species may form immediately as a result of UV exposure, or result from the inflammatory response which often follows UV-induced injury.
  • an inflammatory infiltrate may be evident histopathologically even in the absence of erythema, and may result in continued exposure of the dermis to free radicals, days after the UV- induced damage has occurred (Kligman, A. M., JAMA, 1969, 210:2377-2380; Lavker et al . , J. Am . Acad .
  • transgenic hairless mouse model has now been developed which expresses a full length or truncated human elastin promoter/reporter gene.
  • These transgenic hairless mice express human elastin promoter activity in a tissue-specific and developmentally regulated manner. Not only can quantitative data be obtained in these mice or fibroblasts derived therefrom after only a single exposure to ultraviolet radiation, but these hairless mice can also be used in long term phototoxicity studies.
  • a single exposure to UVB in transgenic hairless mice having a truncated elastin promoter resulted in about a 20 to 30 fold increase in elastin activity as compared to an 8-fold increase reported for prior art models expressing the full length promoter. Accordingly, this transgenic hairless mouse and fibroblasts derived from this hairless mouse are useful as in vivo and in vi tro models to study cutaneous photoaging and in the identification of agents which may protect against photodamage.
  • the present invention also relates to an in vi tro system and method for identifying agents capable of protecting against oxidative damage via a mouse fibroblast culture derived from a transgenic hairless mouse capable of expressing a full length or truncated human elastin promoter and a means for generating reactive oxygen species within the mouse fibroblast cultures .
  • An object of the present invention is to provide a transgenic hairless mouse capable of expressing a full length or truncated human elastin promoter.
  • Another object of the present invention is to provide an in vi tro system for identifying agents capable of inhibiting or preventing oxidative damage comprising a mouse fibroblast culture derived from a transgenic hairless mouse capable of expressing a full length or truncated human elastin promoter and a means for generating reactive oxygen species within the mouse fibroblast culture.
  • Yet another object of the present invention is to provide a method of identifying agents capable of inhibiting or preventing oxidative damage using this in vi tro system.
  • transgenic hairless mouse model has been developed which permits the investigation of human elastin promoter activity in response to ultraviolet irradiation both in vivo by direct irradiation of mouse skin, and in vi tro by irradiation of dermal fibroblasts grown from skin explants of these mice.
  • the hairless mouse used in the production of these transgenic mice is of the strain Crl : SKHl-hrBR (Charles River) as this hairless strain of mice is well characterized and used routinely in preclinical dermatological and photobiological research.
  • the transgenic hairless mice of the present invention are capable of expressing a full length or truncated elastin promoter.
  • truncated human elastin promoter it is meant a human elastin promoter shorter than the full length 5.2 kb human elastin promoter such as pEP62 , pEP35, pEPIO, pEP27, and pEP6 (Kahari et al . , J. Biol . Chem . , 1990, 265 (16) :9485-9490) which is activated by UV.
  • the truncated elastin promoter is pEP6. It is also preferred that the promoter be linked to a reporter gene such as the chloramphenicol acetyltransferase reporter gene (CAT) for ease in detecting activity of the full length or truncated promoter .
  • CAT chloramphenicol acetyltransferase reporter gene
  • mice were first produced by Sams et al . using very large amounts of ultraviolet radiation (J " . Invest . Dermatol . , 1964, 43:467-471) .
  • J " . Invest . Dermatol . , 1964, 43:467-471) was produced by Sams et al . using very large amounts of ultraviolet radiation.
  • one group of mice received 1,040 human minimal erythema doses (MEDs) over 3 months from a bank of fluorescent tubes, while another group received 13,000 MEDs given over 52 weeks in 260 treatments.
  • Elastosis was demonstrated by histochemical staining for elastin and, in irradiated mice, demonstrated an increased elastin staining.
  • mice a dose-response relationship for elastin promoter activity after only a single dose of UV has been observed.
  • a single dose of UVB 491.4 mJ/cm 2
  • UVA 38.2 J/cm 2
  • mice four or five days old must be used since at this age, visible hair growth is not yet present.
  • mice require addition of 8-methoxypsoralen prior to UVA exposure to achieve a significant increase in elastin promoter activity.
  • 8-methoxypsoralen 8 -MOP
  • UVA 8-methoxypsoralen
  • PUVA 8-methoxypsoralen
  • Treatment of skin diseases with PUVA results in clinical alterations in treated skin similar to those observed in chronically photodamaged skin.
  • PUVA-treated patients develop non-melanoma skin cancers, pigmentary alterations and wrinkling characteristics of sun- induced changes.
  • mice of the present invention not only is a quantifiable increase in elastin promoter activity in UVB-treated mice observed after a single dose of UVB, but these mice can also be used in long term phototoxicity studies. Further, a 20 to 30 fold increase in promoter activity was observed in mice of the present invention expressing a truncated human elastin promoter following a single dose of UVB, thus demonstrating that transgenic hairless mice capable of expressing a truncated elastin promoter provide a more sensitive model as compared to prior art mice expressing the full length promoter. With this increase in sensitivity to UV, it is believed that application of psoralen to fibroblasts and/or mice will no longer be required in experiments investigating UVA effects.
  • the present invention also relates to methods of identifying compounds capable of inhibiting cutaneous photodamage with this transgenic hairless mouse model.
  • a test compound is applied to the skin of a transgenic hairless mouse capable of expressing a full length or truncated human elastin promoter.
  • the transgenic hairless mouse is then exposed to ultraviolet radiation, either solar simulating, UVB or UVA, and human elastin promoter activity in the mouse is determined.
  • the human elastin promoter activity is then compared to that in control transgenic hairless mice also exposed to an equivalent dose of ultraviolet radiation which were not treated with the test compound to determine whether or not the test compound provided protection against the ultraviolet radiation.
  • fibroblast cells derived from a transgenic mouse capable of expressing a full length or truncated human elastin promoter are treated with a test compound.
  • the treated fibroblast cells are then exposed to solar simulating, UVB or UVA radiation and human elastin promoter activity in the fibroblast cells is determined. This activity is compared to control fibroblast cells from the transgenic mice exposed to the same dose of solar simulating, UVB or UVA radiation but which were not treated with the test compound to determine if the test compound provided protection against the exposure.
  • Oxidative damage is also believed to play a role in dermal damage from UV radiation.
  • the generation of photoaging clinically evident as wrinkling and sagging of skin, may result more from free radical-induced mechanisms than UV-induced skin cancers which originate in the epidermis.
  • Evidence for this includes the greater sensitivity of fibroblasts to free radical-induced damage as compared to keratinocytes (Applegate, L.A. and Frenk, E. Photodermatol . Photo immunol . Photomed . , 1995, 11:95-101; Moysan et al . , Photodermatol . Photoimmunol . Photomed . , 1995, 11:192-197; Masaki, H. and Sakurai, H. J.
  • UVA-induced photoaging may be the primary means by which UVA- induced photoaging takes place.
  • Test agents suspected of providing protection against oxidative damage can be added to the hairless mouse fibroblast culture prior to addition of the means for generation of reactive oxygen species.
  • the means for generating reactive oxygen species is then added and human elastin promoter activity is determined in the mouse fibroblast culture after a selected time period.
  • the time period for determination of human elastin promoter can be selected in accordance with routine experiments wherein optimum time span for incubation of fibroblasts, derived from the skin of the transgenic mice, with a hypoxanthine and xanthine oxidase system is determined.
  • optimum time span for incubation is determined by exposing cells to a hypoxanthine and xanthine oxidase system for increasing amounts of time, and determining promoter activity at various times throughout a 24 hour incubation. Optimum time is determined as the point at which CAT activity peaks.
  • elastin promoter activity is determined by measuring expression of the reporter gene, i.e. the CAT activity, in the hairless mouse fibroblast culture. Human elastin promoter activity in the hairless mouse fibroblast culture exposed to the test agent is then compared to elastin promoter activity in a control hairless mouse fibroblast culture not exposed to the test agent but still exposed to the means for generating reactive oxygen species. Agents providing protection against oxidative damage are identified as those test agents which decrease human elastin promoter activity in the hairless mouse fibroblast culture exposed to the test agent and the means for generating reactive oxygen species as compared to the control hairless mouse fibroblast culture.
  • Example 1 Transgenic mice expressing the human elastin promoter
  • mice Although phenotypically normal, these mice express the human elastin promoter when assayed for CAT activity (Hsu-Wong et al . , J " . Biol . Chem . , 1994, 269: 18072-18075; Bernstein et al . , J Invest Dermatol , 1995, 105, 269-273; and Bernstein et al . , Photochem Photobiol , 1996, 64:369-74) .
  • Fibroblast cultures are established from the skin of these hairless transgenic mice by explanting tissue specimens onto plastic tissue culture dishes and allowing cells to migrate to the area of the dish surrounding the explants .
  • the primary cultures are maintained in Dulbecco ' s modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum (FCS) , 2 mM L-glutamine and antibiotics, at 37°C. After exposure to ultraviolet radiation, the cells are incubated in DME medium supplemented with 10% fetal calf serum for 24 hours, then harvested for determination of CAT activity as described in Example 4.
  • DMEM Dulbecco ' s modified Eagle's medium
  • FCS fetal calf serum
  • FCS fetal calf serum
  • Example 3 Exposure of fibroblast cultures to free radicals generated by hypoxanthine/xanthine oxidase DMEM with 10% FCS is removed, cells are rinsed in phosphate buffered saline (PBS) , and DMEM is replaced without the addition of FCS. Both 500 ⁇ M hypoxanthine (Sigma Chemical Co., St. Louis, MO) and 80 mU/ml xanthine oxidase (Sigma Chemical Co., St. Louis, MO) are then added to fibroblast cultures and incubated at 37°C for a selected time period. These concentrations of hypoxanthine and xanthine oxidase were selected based on previous work by Mitchell et al .
  • PBS phosphate buffered saline
  • the time for incubation of cells with hypoxanthine/xanthine oxidase can be selected by exposing cells to hypoxanthine/xanthine oxidase for 15, 30, 60, 90 and 120 minutes and determining CAT activity as outlined in Example 4. After exposure to hypoxanthine/xanthine oxidase, cells are rinsed in PBS and incubated in DMEM with FCS for the time in which maximal promoter activation is determined to occur after exposure to hypoxanthine/xanthine oxidase. Control cells are treated in an identical fashion without the addition of hypoxanthine/xanthine oxidase.
  • hypoxanthine/xanthine oxidase In addition to hypoxanthine/xanthine oxidase treatment alone, cells are also treated with hypoxanthine/xanthine oxidase plus 10,000 U/ml of catalase (Sigma Chemical Co., St. Louis, MO) co- incubated for a selected time, and harvested as outlined above. Fibroblasts from the hairless mice representing the same litter are used for any given experiment. Four dishes of cells are used for each experimental condition (control, hypoxanthine/xanthine oxidase, and hypoxanthine/xanthine oxidase+catalase) , and experiments are repeated in duplicate, yielding a total of eight values for each experimental condition.
  • catalase Sigma Chemical Co., St. Louis, MO
  • hypoxanthine/xanthine oxidase and hypoxanthine/xanthine oxidase+catalase are determined using the trypan blue (Sigma Chemical Co., St. Louis, MO) exclusion method (Ausubel et al . , Short Protocols in Molecular Biology, John Wiley & Sons, Inc., 2nd ed., New York, 1992, p.11-24), and a paired t-test analysis is performed for statistical evaluation of the data.
  • Example 4 CAT Assay
  • CAT activity is determined.
  • the specimens are homogenized in 0.25 Tris- HCl, pH 7.5, using a tissue homogenizer (Brinkmann Instruments, Inc., Westbury, NY). The homogenates are centrifuged at 10,000 X g for 15 minutes at 4°C and the protein concentration in the supernatant determined by a commercial protein assay kit (Bio-Rad Laboratories, Richmond, CA) .
  • a closely spaced array of seven Westinghouse FS-40 sunlamps is used which delivers uniform irradiation at a distance of 35 cm. Irradiating with UVA is performed using seven Sylvania FR40T12 PUVA lamps in the above mentioned array, filtered through window glass of 2 mm thickness to remove wavelengths below 320 nm. The energy output at 35 cm is measured with a Solar Light model 3D UVA and UVB detector (Solar Light Company, Philadelphia, PA). The output of FX-40 sunlamps is 23.4 units/hour of UVB at 38 cm, where each unit is equivalent to
  • the output for FR40T12 PUVA lamps filtered through window glass is 2.02 mW/cm 2 , with no detectable UVB radiation.
  • a Multiport Solar Simulator (Solar Light Company, Philadelphia, PA) containing a xenon arc lamp filtered through a Schott WG 320 filter (Schott Glastechnike, Mainz, Germany) is used to administer solar simulating radiation (SSR) .
  • SSR solar simulating radiation
  • the output of the solar simulator is measured by means of a 3D UV meter (Solar Light Company) and displayed as human minimal erythema doses (MEDs) .
  • MEDs minimal erythema doses
  • mice are placed under the center of the light array and restrained with adhesive tape, exposing their dorsal surfaces to the ultraviolet radiation at a distance of 35 cm from the fluorescent tubes.
  • For the Multiport Solar Simulator light guides from the solar simulator are placed in light contact with the dorsal surface of the mice, which are restrained to prevent movement while SSR is administered. Untreated control mice are restrained in a similar manner.
  • Fibroblast cultures as described above are exposed for 5, 10, 20, 40 and 80 seconds of UVB corresponding to doses of 0.7, 1.4, 2.7, 5.5 and 10.9 mJ/cm 2 , respectively.
  • Cultures are exposed to UVA for 2.3, 4.6, 9.2 and 18.4 minutes corresponding to doses of 0.3, 0.6, 1.1 and 2.2 J/cm 2 .
  • tissue culture medium is removed from cells and replaced with a thin layer of phosphate buffered saline (PBS) sufficient to cover the cells. Control unirradiated cells are also placed in PBS. Medium is replaced in all dishes immediately after the last light dose is administered. Only fibroblasts from mice in the same litters are used for any given experiment and utilized in the first few passages. Two dishes of cells are used for each time point.
  • PBS phosphate buffered saline

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Abstract

L'invention a trait à des souris glabres transgéniques qui expriment un promoteur d'élastine humain en pleine longueur ou tronqué, et à des cultures fibroblastiques provenant de ces souris transgéniques. Cette invention concerne également des méthodes d'identification de composés qui peuvent inhiber la photodestruction cutanée de ces souris glabres transgéniques ou des cultures fibroblastiques provenant de ces souris. En outre, ladite invention concerne un système in vitro d'identification des agents qui peuvent inhiber ou prévenir la destruction oxydative dans cette culture fibroblastique, système dans lequel des espèces d'oxygène réactives sont générées au sein de ladite culture, et une méthode d'identification d'agents de prévention de la destruction oxydative.
PCT/US2000/004703 1999-02-24 2000-02-22 Modele in vivo et in vitro pour photovieillissement cutane et destruction oxydative Ceased WO2000050892A1 (fr)

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Application Number Priority Date Filing Date Title
CA002362585A CA2362585A1 (fr) 1999-02-24 2000-02-22 Modele in vivo et in vitro pour photovieillissement cutane et destruction oxydative
EP00911941A EP1163514A4 (fr) 1999-02-24 2000-02-22 Modele in vivo et in vitro pour photovieillissement cutane et destruction oxydative
JP2000601437A JP2002542764A (ja) 1999-02-24 2000-02-22 皮膚の光老化および酸化的損傷のためのインビボおよびインビトロモデル
AU33754/00A AU761728B2 (en) 1999-02-24 2000-02-22 An (in vivo) and (in vitro) model for cutaneous photoaging and oxidative damage

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US12026999P 1999-02-24 1999-02-24
US60/120,269 1999-02-24

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CN116745417A (zh) 2021-03-09 2023-09-12 优志旺电机株式会社 光衰老细胞的检测或定量方法、以及其应用、光衰老细胞的制作方法

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WO1996037237A1 (fr) * 1995-05-24 1996-11-28 Thomas Jefferson University Modele in vivo et in vitro de photovieillissement cutane

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JP2974446B2 (ja) * 1991-04-30 1999-11-10 中外製薬株式会社 脱毛症マウス

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Publication number Priority date Publication date Assignee Title
WO1996037237A1 (fr) * 1995-05-24 1996-11-28 Thomas Jefferson University Modele in vivo et in vitro de photovieillissement cutane

Non-Patent Citations (5)

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Title
BERNSTEIN ET AL.: "Ultraviolet Radiation Activates the Human Elastin Promoter in Transgenic Mice: A Novel In Vivo and In Vitro Model of Cutaneous Photoaging", J. OF INVEST. DERMATOL.,, vol. 105, no. 2, August 1995 (1995-08-01), pages 269 - 273, XP002927947 *
KATCHMAN ET AL.: "Transforming Growth Factor-beta Up-Regulates Human Elastin Promoter Activity in Transgenic Mice", BIOCHEM. BIOPHYS. RES. COMM.,, vol. 203, no. 1, 30 August 1994 (1994-08-30), pages 485 - 490, XP002927948 *
LEDO ET AL.: "Glucocorticosteriods Up-Regulate Human Elastin Gene Promoter Activity in Transgenic Mice", J. INVEST. DERMATOL.,, vol. 103, no. 5, November 1994 (1994-11-01), pages 632 - 636, XP002927949 *
See also references of EP1163514A4 *
UITTO ET AL.: "Molecular Aspects of Photoaging", EUR. J. DERMATOL.,, vol. 7, no. 3, 1997, pages 210 - 214, XP002927950 *

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JP2002542764A (ja) 2002-12-17
AU3375400A (en) 2000-09-14
AU761728B2 (en) 2003-06-05
CA2362585A1 (fr) 2000-08-31
EP1163514A4 (fr) 2002-09-18
EP1163514A1 (fr) 2001-12-19

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