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