METHOD FOR OBTAINING A SPRAY-ON CELLULAR COMPOUND OF HUMAN FIBROBLASTS AND KERATINOCYTES IN SOLUTION AND THE USE THEREOF AS A REGENERATIVE AGENT IN SKIN LESIONS

摘要

The present invention relates to a process for obtaining a compound of cellular spraying for fibroblasts and human keratinocytes and its use as regenerative agent of cutaneous lesions, mainly in diabetic ulcers, first, second, and third grade burns, and a substitute for the use of flaps techniques. The process for obtaining a compound of cellular spraying for fibroblasts and human keratinocytes includes the steps of separating the dermis and epidermis layers, incubation of the epidermis layer, disintegration of dermis layer and proliferation of fibroblasts, disintegration of the epidermis layer, proliferation of keratinocytes in suspension, preparation of the cross-linking reaction agent solution of the cellular spraying solution, and preparation of cellular spraying solution. The compound for cellular spraying of fibroblasts and human keratinocytes increases the application efficiency and ease of it, in order to be an economic alternative, fast, and easy to use.

主权项

1. A process for obtaining a compound of cellular spraying for fibroblasts and keratinocytes comprising the steps of:
a) separation of a dermis layer and an epidermis layer; b) incubating of the epidermis layer to obtain a trypsinized epidermis; c) disintegrating the dermis layer and proliferating fibroblasts; d) disaggregating the epidermis layer; e) proliferating keratinocytes in suspension; f) preparing a cross-linking reaction agent solution of the cellular spraying solution; and g) preparing of cellular spraying solution wherein in the step a) a sample of skin is obtained from a patient under sterile conditions by using a dermatome to obtain a skin sheet; washing the skin sheet at least 3 times with a saline solution of phosphates (PBS) 1× containing 10% v/v of antibiotic/antimiotic; washing at least 3 times with a PBS 1× solution containing 1% v/v of antibiotic/antimiotic; incubating with a cellular disintegration proteolytic enzyme at a concentration of 1.79 units/mg at a temperature of 4° C. for at least 16 hours to get the epidermis layer and the dermis layer; after 16 hours, separating the two layers using sterile tweezers and placing the layers in separate and sterile containers.