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Regenerative medicine is a modern approach of dermatological treatment, using Epidermal Cells of the interfollicular epidermis (ESC) for their effect in skin regeneration in chronic ulcers and burns, melanoma, vitiligo, junctional epidermolysisbullosa. Intraepidermal injections of autologous epidermal cell suspension can be a new and very promising treatment for mano other cutaneous disorders as non scarring alopecia (alopecia areata, androgenic alopecia) or scarring alopecia (Lichern Plano Pilaris alopecia, Discoid Lupus Erithematosus alopecia), anti-aging therapies. The intraepidermal injection of an autologous epidermal cell suspension is a simple, fast and safe surgical procedure: a small, thin portion of the epidermis of the patient undergoes a treatment where a suspension with all the cells collected from the epidermis is injected into the skin. The epidermal grafts were then incubated in a trypsin solution (Trypsyn 0.5 g/ EDTA 0.2 solution, Sigma-Aldrich co) for 45 minutes at 37oC (Plasmatherm Barkley). After incubation, the trypsin solution was discarded and the tissues were washed with HBSS. The epidermis (thin yellow layer) was treated with a scalpel blade to separate the cells. The supernatant was suctioned through a sterile syringe and then cultured in Dulbecco's modified Eagle's medium (DMEM; Sigma Aldrich, Saint Louis, USA) supplemented with 10% (v/v) heat-inactivated foetal bovine serum (EuroClone, Devon, UK) and 1% penicillin/streptomycin solution (Sigma Aldrich) at 37 oC in a humidified atmosphere of 95% air and 5% CO2. The cells were seeding in a 25oC at day 0, the cells showed a heterogeneous appearance with regard to shape and size and their nucleus was almost detectable. At day 7, the cells maintained a heterogeneous appearance with regard to shape and size, even though more spheroidal elements were observed. Cells were generally in suspension, only some microspots of fibroblastoid-shaped cells were well attached to the flask surface m2 cell culture flask at a density of about 20000 cells/cm2. Our preliminary study show that an suspension contains a significant number of viable cells (59,40%, SD ± 6.07%) that survive at day 7 in culture (63.55% , SD ± 5.41%). The number of epidermal cells in each sample is significant as keratinocytes (in greater quantity) and melanocytes (1900 ±178 melanocytes/mm2 as reported by Khodadadi L) (6) are detected. Our research is currently continuing and it is focusing on the typing of the different cells in the suspension and evaluating the presence and the nature of stem cells.

Intraepidermal Injections of Autologous Epidermal Cell Suspension: A new promising approach to Dermatological Disorders. Preliminary Study