In vitro skin equivalent, useful e.g. for evaluating skin pigmentation phenomena and in tanning test, comprises epidermis equivalent comprising keratinocyte forming basal layer, superficial layer and dermis equivalent, and melanocytes

摘要

In vitro skin equivalent comprises: epidermis equivalent comprising keratinocyte forming at least one basal layer, at least one superficial layer and at least one dermis equivalent; and melanocytes producing constitutive melanin and living fibroblasts, where the entire melanocytes are in the basal layer of the epidermis equivalent. Independent claims are included for: (1) a process for preparing the skin equivalent, comprising (a) contacting fibroblasts and collagen solution and then incubated for a sufficient time to obtain a collagen matrix which are contacted in which the fibroblasts are distributed, constituting a dermis equivalent, (b) seeding the dermis equivalent by a mixture of keratinocytes and melanocytes on dermis equivalent obtained in (a) and culturing by immersing in a liquid medium, and (c) emerging keratinocytes and melanocytes from the culture on the dermis equivalent obtained in (b) and culturing at the air-liquid interface to obtain a multilayered epidermis equivalent containing melanocytes, on dermis equivalent containing fibroblasts in a collagen matrix constituting skin equivalent; and (2) a process for evaluating the ability of an agent to modulate the pigmentation, comprising (a1) applying the agent to the in vitro skin equivalent, and (b1) comparing the pigmentation (i) of the in vitro skin equivalent to which the evaluating agent is applied, with pigmentation (ii) of a control skin equivalent not subjected to the agent.