Neural Stem Cell Harvest and Culture using the High Speed Centrifugation from Rat Brain / 대한마취과학회지

ABSTRACT

BACKGROUND: During recent two decades of crucial revision of some cornerstone concepts has opened new horizons in neurosciences. Modern basic viewpoints include the idea of high CNS plasticity which means not only rearrangement of neurons and their interconnections, but also the formation of new neural cells in humans and animals during their whole life span. The purpose of this study is to harvest neural stem cell from the adult rat brain using the high speed centrifugation method and study the characteristics of these cell. METHODS: 60 rats (Fisher 344, 150-160 g) brain were saved under inhalation anesthesia and dissect the subventricular zone under the microscope. The brain tissue was digested with enzyme to make a cell suspension. The cell suspension was processed high speed centrifugation to separate the neural stem/progenitor cells according to the buoyancy. After 2 weeks culture, immuno-staining (O4, GFAP, Nestin, beta-tubulin III and DAPI) were performed and replated the cultured cells. RESULTS: The 2 weeks culture cells were positive 92.8% in Nestin, 91.5% in O4 and 87.6% in Gal-C. But only positive 1.4% in beta-tubulin III and 5.5% in GFAP. And replated cell culture shows similar results compared to the primary culture. CONCLUSIONS: With this high speed centrifugation method, authors can harvest neural stem/progenitor cells from the adult rat brain. Although we have many limitations using these cell in clinical trial, but we can afford to next step on neural stem cell research.