Bone marrow mononuclear cells characterization and participation in the Wallerian degeneration process.

USACH, V; GOITIA, B; SETTON-AVRUJ CP

Buzios, Brasil

Congreso; I Congreso IBRO/LARC de Neurociencias de América Latina, Caribe y Península Ibérica, Buzios, Brasil; 2008

IBRO/LARC

Bone marrow mononuclear cells characterization and participation in the Wallerian degeneration process Vanina Usach, Belén Goitia and Patricia Setton-Avruj Cátedra de Química Biológica Patológica, FFyB, IQUFIB, UBA-CONICET, IIMHNO-UBA We have previously described the spontaneous migration or after an intraortically injection of fresh bone marrow mononuclear cells (BMMC) CD34+ to the distal stump of a ligated sciatic nerve, an irreversible model of Wallerian degeneration (WD). The aim of the present work is to demonstrate in a reversible model of Wallerian degeneration such as the crush of the sciatic nerve whether BMMCs migrate to the distal area in order to participate in the degeneration  - regeneration process, and to characterize the BMMCs and the mesenchymal stem cells in order to know which is the population that migrate to the injured nerve during the WD process. Adult Wistar rats were submitted to the crush of the right sciatic nerve and were sacrificed at different survival times. The nerve was dissected into proximal and distal areas; the left sciatic nerve, contralateral nerve, was taken as a control. BMMC were isolated from the bone marrow extruded from tibia and femur bones. A group was immediately processed and the other was seeded. After 24 and 48hs the non-adherent cells were separated and the adherent cells were grown to confluence. Both groups were analyzed by Western blot. Fresh isolated or cultured BMMC were dyed with a fluorescent probe and injected intravenously immediately after crushing the sciatic nerve in order to evaluate the migration of these cells to the injured area. The characterisation of cultured BMMCs shows a heterogeneous population of small-rounded, spindle-shaped and large-flattened morphology; they showed fibroblast-like morphology at reaching confluence. After 24 and 48hs in culture the adherent and non-adherent cells are CD34+, while adherent cells are CD45-, CD11b-, vimentin+, Thy 1.1+. We demonstrated the migration of fresh isolated BMMC to the crushed area and to the dital area of the sciatic nerve during the first week post injury. Our results demonstrate the existence of a progenitor population in BMMC that spontaneously migrates to the injured nerve to participate in the degeneration-regeneration process.