Flow Cytometric Analysis of Cultured Adult Neural Stem Cells from Guinea Pig Spiral Ganglion / 대한이비인후과학회지

ABSTRACT

BACKGROUND AND OBJECTIVES: To make stem cell therapy successful as one of treatment options for sensorineural hearing loss, it is essential to culture and obtain sufficient amounts of adult neural stem cells, as well as separating them from adult auditory organs. This study was designed to investigate the proportion of cultured adult neural stem cells and its differentiated cells from guinea pig spiral ganglion. MATERIALS AND METHOD: The spiral ganglions from guinea pigs of 3-6 month age were obtained. The tissues were digested with 0.25 % trypsin and 10 mg/mL of DNase I, cells were then cultured with neurobasal medium (DMEM/F12 containing B27 supplement, L-glutamin, gentamycin) and added with 20 ng/mL of epidermal growth factor and 10 ng/mL of fibroblast growth factor. After 3 passages of culture, neural stem cells and differentiated cells were analyzed with the flow cytometric method. RESULTS: We concluded that neural stem cells were successfully cultured from spiral ganglions and these cells were in process of differentiation into neurons and Schwann cells. The results of flow cytometric analysis of cells in culture medium showed that 1.7% of cells (cell count of 24,300) expressed nestin, 3.45% polysialylated neural cell adhesion molecule, 7.19% (cell count of 66,300) neural cell adhesion molecule, and 3.57% beta III tubulin. CONCLUSION: Though obtaining adult neural stem cells from adult spiral ganglion was successful, the cell count was small. Further studies on the subject of making proper culture medium are needed to obtain adequate amounts of adult neural stem cells.