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Artificial epidermis in-vitro model for detecting sensitization of material and screening medicament

An allergenic and artificial technology, applied in the field of biomedicine, can solve the problems of lack of stratum corneum barrier, can not be used to detect insoluble substances and sensitization of external preparations, can not reflect the transdermal performance of external immunomodulatory drugs, etc., to overcome Not easy to industrialize, easy to control quality, and small batch-to-batch difference

Active Publication Date: 2013-01-23
CHUGOKU IGAKU KAGAKUIN HIFUBIYOU KENKYUSHO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, the current models established with monocytic cells are all single-cell two-dimensional culture models, lacking the stratum corneum barrier, so they can only be used to detect the sensitization of soluble substances, and cannot be used to detect the sensitization of insoluble substances and external preparations. sensitivity, and cannot reflect the transdermal performance of topical immunomodulatory drugs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0027] The preparation of human keratinocytes refers to the method in the following literature: Boyce ST, Ham RG. Calcium-regulated differentiation of normal human epidermal keratinocytes in chemically defined clonal culture and serum-free serial culture. JInvest Dermatol, 1983, 81 (1Suppl): 33s-40s..

[0028] The preparation of human fibroblasts refers to the method in the following literature: Gray BA, Gollin SM. Rapid cell culture procedure for tissue samples. Am J Med Genet, 1987, 28(2): 521-6.

Embodiment 1

[0030] 1. The in vitro model of artificial epidermis used for detection of substance sensitization and drug screening in this example is obtained by co-cultivating artificial epidermis and THP-1 cell line cells.

[0031] 2. In the present embodiment, the construction method of the artificial epidermis in vitro model comprises the following steps:

[0032] 1) Take the suspension of human keratinocytes and make 5×10 5 The density of / ml was inoculated into the upper chamber of the inserted culture dish. Before inoculation, the lower chamber of the inserted culture dish was filled with human fibroblasts. 2 Cultured in an incubator for 3 days, after the cells grew to a confluent state, transferred to the air-liquid interface for 14 days to obtain an artificial epidermis;

[0033] 2) THP-1 cell line cells were mixed with 5×10 5 Inoculate in a well plate at a density of / ml, transfer the obtained artificial epidermis together with the upper chamber of the insert culture dish to th...

Embodiment 2

[0045] 1. The in vitro model of artificial epidermis used for detection of substance sensitization and drug screening in this example is obtained by co-culturing artificial epidermis and MUTZ-3 cell line cells.

[0046] 2. In the present embodiment, the construction method of the artificial epidermis in vitro model comprises the following steps:

[0047] 1) Take the suspension of human keratinocytes and make 2×10 5 The density of / ml was inoculated into the upper chamber of the inserted culture dish. Before inoculation, the lower chamber of the inserted culture dish was filled with human fibroblasts. 2 Cultured in an incubator for 5 days, after the cells grew to a confluent state, they were transferred to an air-liquid interface for 14 days to obtain an artificial epidermis;

[0048] 2) MUTZ-3 cell line cells were mixed with 2×10 5 The density per ml was inoculated in the well plate, and the medium used was MEM medium containing 20% ​​FBS and 10% conditioned medium of feeder...

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Abstract

The invention provides an artificial epidermis in-vitro model for detecting the sensitization of a material and screening an external skin immune regulation medicament or pharmacodynamics evaluation. The model is prepared by culturing artificial epidermis and monocytic series cells together. The artificial epidermis in-vitro model has the following advantages that: 1, the sensitization of variouscompounds, external skin preparations and cosmetics can be evaluated by detecting the change of related cell factors and cell surface markers in the model; when the sensitization of the material is judged, the model has high sensitiveness, high specificity and high accuracy, the detection method is simple and the model has a wide application range; 2, the model can be used for screening the external skin immune regulation medicament or pharmacodynamics evaluation; 3, the construction cost of the model is low, bath-to-batch variation is small, and quality is controlled easily; and 4, the modelhas the readily available materials and can be conveniently produced on scales.

Description

technical field [0001] The invention relates to an artificial epidermis in vitro model, in particular to an artificial epidermis in vitro model for detecting substance sensitization and for screening or pharmacodynamic evaluation of immunomodulatory drugs externally applied to the skin, belonging to the technical field of biomedicine . Background technique [0002] In recent years, with the aggravation of environmental pollution, the abuse of chemicals, and the increase of external harmful substances, the incidence of skin allergies has been increasing year by year. The evaluation of the allergenicity of substances has attracted more and more attention. The traditional evaluation method is mainly based on animal experiments. However, under the influence of the global animal protection movement, developed countries have generally carried out substitution, reduction and optimization as the core In the "3R" movement, in vitro alternative tests will gradually replace animal tes...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N5/0786C12Q1/02G01N15/10G01N33/53
Inventor 马鹏程刘维达曹玉萍周武庆
Owner CHUGOKU IGAKU KAGAKUIN HIFUBIYOU KENKYUSHO