Endoneural fibroblasts isolation and culture

• Main Authors: Leslie Leal, Sandra Perdomo, Clara Spinel
• Format: Article
• Language: English
• Published: Universidad Nacional de Colombia 2004-07-01
• Series: Acta Biológica Colombiana
• Subjects: Nervio Ciático; Fibroblastos Endoneurales; Cultivo en Monocapa
• Online Access: https://revistas.unal.edu.co/index.php/actabiol/article/view/27105

Fibroblasts which are tissue-specific, constantly degrade and synthesize the different elements of the extra-cellular matrix (ECM), while at the same time remodel tissues that are being repaired. Dermal fibroblasts are well studied both in vitro and in vivo, and are used to regenerate dermal EMC which in turn supports the regeneration of the epidermis. Confluence of dermal or periodontal fibroblasts takes place between 8 and 10 days of culture. In the process of regeneration of damaged peripheral nerves, Schwann's cells secrete neurotrophic and neurotropic growth factors and some of the EMC elements needed for regeneration to take place, which makes them the most studied and used cells in culture. So far, endoneural fibroblasts (EF) have not been considered as important elements in nerve regeneration, mainly because they may occasionally form fibromes that hinder regeneration. But there is evidence that they may play a role in the remodeling of the EMC, through the secretion of metalloproteins that modify the pre-Nerve Growth Factor (preNGF) secreted by Schwann's cells into active NGF, which promotes neurites regeneration. The aim of this study was is to isolate EF from sciatic nerves taken from mature rats, and to obtain them in purified culture. A number of methods of dissection and digestion were developed to obtain primary pure EF cultures as well as to study them in the way Schwann's cells have been studied. Selective isolation of EF was accomplished, reaching confluence between the fourth and the fifth day in monolayer primary culture. Producing a population of EF will make it possible to carry out studies in tridimensional culture and in prosthesis in order to define and develop new alternatives for the regeneration of peripheral nerves.