Isolation of oligodendrocyte precursors from umbilical cord blood for the repair of spinal cord injury

Abstract

Abstract Introduction: Optimizing remyelination through direct delivery of myelin producing cells may offer alternative approaches to restoring neurologic function in patients who sustain brain and spinal cord injuries. We hypothesized that umbilical cord blood (UCB) contains cells capable of differentiating into precursors of neural lineages, specifically oligodendrocytes, which could be used for cellular repair of neural tissues in patients who have suffered injury. Methods: Mononuclear cells were isolated from UCB and cultured in media containing platelet-derived growth factor (PDGF) (5ng/mL), neurotrophin-3 (1ng/mL), tri-iodothyronine (30ng/mL), vascular endothelial growth factor (VEGF) (10ng/mL), and 10% FBS. An adherent cell population was observed after one week. Cells were fixed and stained using immunocytochemical techniques for hematopoietic markers CD45 and CD34, human leukocyte antigen (HLA) class I and II, microglial markers CD68 and CD11b, astrocyte marker GFAP, oligodendrocyte markers O1, O4, and myelin basic protein (MBP), and neuronal markers NeuN and B-tubulin III. RT-PCR was performed using primers specific to oligodendrocyte precursors: MBP and 2′,3′,-cyclic nucleotide 3′-phosphodiesterase (CNPase). Results: We observed that UCB contains cells capable of differentiation along the oligodendrocyte lineage, evidenced by staining for CD45, O1, O4, MBP, HLA I, and HLA II. Expression of MBP and CNPase was demonstrated by RT-PCR. Conclusions: The pattern of RNA production and protein expression observed in UCB-derived cells suggests that they may be capable of oligodendrocyte differentiation. These cells may have the potential for therapeutic applications in patients with diseases or injuries of the central and peripheral nervous system.