Methods for producing hair microfilms and de novo papillae and their use for in vitro assays and in vivo implants

摘要

De novo papilla comprising a cellular aggregate of mammalian dermal hair papilla (DPF) fibroblasts, the cellular aggregate exhibiting the shape and size of a physiological dermal papilla (DP) of a hair follicle after its isolation and after its coating with extracellular matrix proteins, preferably collagen IV, fibronectin and / or laminin, where de novo papilla is produced by a method comprising the steps of: (a) providing at least one dermal papilla (DP) of at least one hair follicle of mammal, (b) isolate dermal papillary fibroblasts (DPF) from the DP by mechanically fixing said DP on the surface of a cell culture vessel, according to which the base sheet is perforated to allow said DPF to migrate , (c) extending the isolated DPFs in monolayer culture without collagen coating, in which said DPFs are passed at least once, (d) condensing the extended DPFs into a cell aggregate that they have the size and shape of the physiological DP, in which said DPFs differ in non-adhesive culture vessels in a concentration of cells per surface of the container from 1,000 to 100,000 DPF / cm2, and (e) coating the de novo papilla with extracellular matrix proteins, preferably collagen IV, fibronectin and / or laminin.