기니아피그 와우신경절에서 배양된 성체 신경줄기세포의 유세포분석법을 이용한 분석 (Flow Cytometric Analysis of Cultured Adult Neural Stem Cells from Guinea Pig Spiral Ganglion)

서지정보

· 발행기관 : 대한이비인후과학회
· 수록지 정보 : 대한이비인후-두경부외과학회지 / 52권 / 2호 / 129 ~ 133페이지
· 저자명 : 김영하, 김창현, 박경호, 박시내, 최미영, 홍옥기, 김지홍, 조상재, 김정현, 장기홍, 여상원

초록

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% β 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.

영어초록

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% β 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.

참고자료

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