Cell culture system for determinging the sensitizing, allergenic and/or irritating effect of a substance

Abstract

The present invention refers to a cell culture system especially for investigating the sensitizing, allergenic and/or irritating effect of substances, comprising a first and a second compartment that can communicate with each other via a permeable interlayer, whereby the first compartment has an epidermis model and the second a cell culture based on immune cells.

1 . A cell culture system for determining the sensitizing, allergenic or irritating effect of one or more substances, comprising:
 a) a first compartment comprising an epidermis model; and   b) a second compartment comprising a cell culture based on immune cells,   wherein the first and second compartments are separated by a permeable interlayer, and wherein an allergen or irritating agent introduced into the first compartment activates the immune cells of the second compartment.   
     
     
         2 . The cell culture system of  claim 1 , wherein the immune cells are antigen presenting cells. 
     
     
         3 . The cell culture system of  claim 2 , wherein the antigen presenting cells are macrophages or dendritic cells. 
     
     
         4 . The cell culture system of  claim 1 , wherein the immune cells are lymphocytes. 
     
     
         5 . The cell culture system of  claim 4 , wherein the lymphocytes are B lymphocytes or T lymphocytes. 
     
     
         6 . The cell culture system of  claim 1 , wherein the immune cells show matching tissue compatibility antigens. 
     
     
         7 . The cell culture system of  claim 1 , wherein the cell culture is substantially free of tissue cells. 
     
     
         8 . The cell culture system of  claim 1 , wherein the immune cells are derived from a human. 
     
     
         9 . The cell culture system of  claim 1 , wherein the immune cells are isolated from blood. 
     
     
         10 . The cell culture system of  claim 1 , wherein the epidermis model has a layered structure. 
     
     
         11 . The cell culture system of  claim 10 , wherein the epidermis model is multilayered. 
     
     
         12 . The cell culture system of  claim 1 , wherein the epidermis model comprises at least one epidermis layer selected from the group consisting of: stratum corneum, stratum lucidum, stratum granulosum, stratum spinosum and stratum basale. 
     
     
         13 . The cell culture system of  claim 1 , wherein the epidermis model comprises live epidermis cells. 
     
     
         14 . The cell culture system of  claim 1 , wherein the live epidermis cells are keratinocytes. 
     
     
         15 . The cell culture system of  claim 1 , wherein the epidermis model is substantially free of immune cells. 
     
     
         16 . The cell culture system of  claim 1 , wherein the epidermis model is an animal epidermis model. 
     
     
         17 . The cell culture system of  claim 16 , wherein the animal epidermis model is a human epidermis model. 
     
     
         18 . The cell culture system of  claim 1 , wherein the one or more substances comprise at least one cosmetic agent. 
     
     
         19 . The cell culture system of  claim 1 , wherein the one or more substances comprise at least one pharmacologically active medicinal agent. 
     
     
         20 . The cell culture system of  claim 1 , wherein the second compartment is filled with an aqueous medium. 
     
     
         21 . A method of using the cell culture system of  claim 1  to examine the sensitizing, allergenic or irritating effect of one or more substances, comprising contacting the first compartment with the substance and detecting an allergenic response in the second compartment. 
     
     
         22 . A method for examining the sensitizing, allergenic and/or irritating effect of an agent, comprising:
 a) contacting the surface of an epidermis model contained in a first compartment with the agent; and   b) determining the activity of a cell culture of immune cells contained in a second compartment, wherein the first compartment and second compartment are separated by a permeable interlayer,   wherein the presence of an activated immune cell in the second compartment indicates that the agent is an allergen or irritating agent.   
     
     
         23 . The method of  claim 22 , wherein the immune cells are isolated from blood. 
     
     
         24 . The method of  claim 22 , wherein the cell culture comprises antigen presenting cells and lymphocytes. 
     
     
         25 . The method of  claim 24 , wherein the antigen presenting cells are derived from monocytes under differentiating conditions. 
     
     
         26 . The method of  claim 22 , wherein the activity of the immune cells is determined by examining the supernatant of the cell culture. 
     
     
         27 . The method of  claim 22 , wherein the activity of the immune cells is determined by assaying for substances produced by activated immune cells. 
     
     
         28 . The method of  claim 27 , wherein the substances produced by activated immune cells are selected from the group consisting of: signal transducers, receptors, messengers, enzymes and antibodies. 
     
     
         29 . The method of  claim 22 , wherein the activity of the immune cells is determined by assaying for messenger substances from keratinocytes of the epidermis model. 
     
     
         30 . The method of  claim 22 , wherein the activity of the immune cells is determined by identifying proliferation of the immune cells. 
     
     
         31 . The method of  claim 22 , wherein the activity of the immune cells is determined by measuring surface markers of the immune cells. 
     
     
         32 . The method of  claim 22 , wherein the activity of the immune cells is determined by measuring signal transduction components of the immune cells. 
     
     
         33 . The method of  claim 22 , wherein the activity of the immune cells is determined by measuring intracellular cytokines of the immune cells. 
     
     
         34 . The method of  claim 22 , wherein the activity of the immune cells is determined by analyzing messenger RNA levels within the immune cells.