In vitro culture and characterization of oligodendrocyte precursor cells derived from neonatal rats.

OBJECTIVES

The pathologic changes of demyelination after spinal cord injury (SCI) significantly impair functional recovery of lesioned spinal cord. At present, transplantation of myelinating cells is regarded as a promising strategy for treating demyelination following SCI. Hence, the In vitro culture and growth, differentiation and proliferation of oligodendrocyte precursor cells (OPCs) were intensively investigated in this study.

METHODS

In vitro cells from cerebral cortices of neonatal rats were primarily cultured and OPCs were then separated by shaking process and differential adhesion. Following cultured in the conditional medium, growth pattern and differentiation of OPCs were continuously studied by both light microscopy and scanning electron microscopy. Furthermore, maturation of OPCs was detected immunochemically and proliferative ability of OPCs In vitro was also evaluated by methyl thiazolyl tetrazolium (MTT) assay.

RESULTS

The distinct stratification of glial cells usually developed around 9-10 days in the primary culture. The OPCs were found primarily living on the surface of confluent astrocytes and these cells typically displayed the simple appearance of immature cells. Furthermore, the OPCs progressively developed in the conditional medium, and these differentiated cells underwent dramatic changes of morphology and also expressed different specific markers. Moreover, the OPCs also proved by MTT assay to proliferate significantly while cultured In vitro.

DISCUSSION

Demyelination prevents recovery of neural function following SCI. Demyelination has already become a potential therapeutic target for this insidious and challenging problem. The In vitro culture and biological characteristics of OPCs are fundamental and necessary for further investigation of cell transplantation in vivo. Growth pattern, differentiation and proliferation are very vital for therapeutical effects of OPCs following transplantation after SCI.