Generation of Demyelination through Use of M. leprae-specific phenolic glycolipid-1 (PGL-1)

Abstract

For myelination, Schwann cells and neuron cells from dorsal root ganglion (DRG) of rat embryos (E16) were cultured in vitro system. The purified DRG cells with anti-mitotic agents and purified Schwann cells were cocultured and then accomplished myelination processing. Treatment of M. leprae-specific phenolic glycolipid-1 (PGL-1) into this coculture system was performed and then accomplished demyelination. Therefore, we identified demyelination processing using antibody of myelin basic protein (MBP). The study of Schwann cell, Neuronal cell, and myelination has been facilitated by the availability to isolate and establish pure population of primary Schwann cells. Moreover, mice serve as an important model for the study of Schwann cell research. The specialized source of neurons from nonneuronal cells were provided in dorsal root ganglia. 1 Adult mammalian DRG neuron cells can survive and regenerate in culture. 2,3,4 There are several researches on purified populations of these neurons. Coculture of purified DRG neurons and Schwann cells can be used in myelin formation. The most widely used method for preparing primary Schwann cell culture uses DRG as the primary source of Schwann cells. The procedure is very simple and produces a highly purified population of Schwann cells in a short time. The method has also been used to prepare Schwann cells from mouse embryos. In this study, we performed a purified population of myelination by coculture of DRG neuronal cells and Schwann cells. The purified DRG cells with anti-mitotic agents and purified Schwann cells were cocultured and then accomplished myelination processing. Treatment of M. leprae-specific phenolic glycolipid-1 (PGL-1) into this coculture system was performed and then accomplished demyelination. Therefore, we identified demyelination processing using antibody of myelin basic protein (MBP). Cultures were incubated at 37 ℃, with 5% CO2. 24 h later, 150 mL of NGF stock solution (40 mM of 5-fluorodeoxyuridine and uridine, 1 mM Arabinofur-anosyl Cytidine (Ara C, Sigma-Aldich, Saint Louis, MO) in NG medium) was added into each well to make the final concentration of 5-fluorodeoxyuridine/uridine 20 mM. Cultures were incubated at 37 ℃ in a 5% CO2 incubator. 5 After 72 hours, 2/3 of the NGF stock solution was changed to NG medium and the cultures were re-fed every 2 days with NG medium. After three