TY - JOUR
T1 - Culture and differentiation of cynomolgus monkey neural stem cells
AU - Yan, Sun Xing
AU - Liu, Xiao Ming
AU - Xiang, Peng
AU - Wang, Peng
AU - Wang, Ding
PY - 2009/11/5
Y1 - 2009/11/5
N2 - BACKGROUND: Studies regarding neural stem cells (NSCs) mainly focus on rodents and human, few of which addressing non-human primate animals. OBJECTIVE: To explore a culture system of cynomolgus monkey NSCs (cmNSCs), additionally, to observe its differential potential. DESIGN, TIME AND SETTING: An in vitro cytology observation was performed at the Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University between July 2008 and April 2009. MATERIALS: Spontaneously aborted cynomolgus monkey fetus with 3-5 months old (gestational age), was provided by Landao Biotechnology Co., Ltd. METHODS: The subventricular zone and the hippocampus were dissected aseptically from the brain of spontaneously aborted cynomolgus monkey fetus, then triturated with fire-polished glass Pasteur pipettes to single cells, and cultured in 25 cm2 culture flask with concentration of (2.0-5.0)× 105. The neurospheres were transferred into polyornithine and fibronectin precoated cell culture dish after 2 weeks when they formed. Each 20 μg/L of epithelium growth factor (EGF) and basic fibroblast growth factor (bFGF) was daily added. The adhesive cells were passaged when they reached about 90% confluency. MAIN OUTCOME MEASURES: The NSCs and its differential potential were identified by surface marker nestin and immunocytochemistry. RESULTS: The neurospheres formed after 1 week culture, and grew with time prolonged. The culture system was changed into adherent culture at 4th week. At 24 hours after plated on the poly-ornithine/fibronectin pre-coated dish, neurospheres adhered to the dish and many cells migrated out of neuroshperes and proliferated as monolayer. The immunocytochemistry showed that NSCs which culture medium contain EGF and bFGF preserve nestin positive, and the cmNSCs that without EGF and bFGF differentiated into astrocytes (GFAP positive), neurons (Tuj-1 positive) and oligodendrocytes (04 positive). CONCLUSION: The culture system which combines suspension culture and adherent culture is fit for the culture of cmNSCs. The cultured cmNSCs has potential to differentiate into astrocytes, neuronal cells and oligodendrocytes.
AB - BACKGROUND: Studies regarding neural stem cells (NSCs) mainly focus on rodents and human, few of which addressing non-human primate animals. OBJECTIVE: To explore a culture system of cynomolgus monkey NSCs (cmNSCs), additionally, to observe its differential potential. DESIGN, TIME AND SETTING: An in vitro cytology observation was performed at the Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University between July 2008 and April 2009. MATERIALS: Spontaneously aborted cynomolgus monkey fetus with 3-5 months old (gestational age), was provided by Landao Biotechnology Co., Ltd. METHODS: The subventricular zone and the hippocampus were dissected aseptically from the brain of spontaneously aborted cynomolgus monkey fetus, then triturated with fire-polished glass Pasteur pipettes to single cells, and cultured in 25 cm2 culture flask with concentration of (2.0-5.0)× 105. The neurospheres were transferred into polyornithine and fibronectin precoated cell culture dish after 2 weeks when they formed. Each 20 μg/L of epithelium growth factor (EGF) and basic fibroblast growth factor (bFGF) was daily added. The adhesive cells were passaged when they reached about 90% confluency. MAIN OUTCOME MEASURES: The NSCs and its differential potential were identified by surface marker nestin and immunocytochemistry. RESULTS: The neurospheres formed after 1 week culture, and grew with time prolonged. The culture system was changed into adherent culture at 4th week. At 24 hours after plated on the poly-ornithine/fibronectin pre-coated dish, neurospheres adhered to the dish and many cells migrated out of neuroshperes and proliferated as monolayer. The immunocytochemistry showed that NSCs which culture medium contain EGF and bFGF preserve nestin positive, and the cmNSCs that without EGF and bFGF differentiated into astrocytes (GFAP positive), neurons (Tuj-1 positive) and oligodendrocytes (04 positive). CONCLUSION: The culture system which combines suspension culture and adherent culture is fit for the culture of cmNSCs. The cultured cmNSCs has potential to differentiate into astrocytes, neuronal cells and oligodendrocytes.
UR - http://www.scopus.com/inward/record.url?scp=77953763743&partnerID=8YFLogxK
U2 - 10.3969/j.issn.1673-8225.2009.45.003
DO - 10.3969/j.issn.1673-8225.2009.45.003
M3 - Article
AN - SCOPUS:77953763743
SN - 1673-8225
VL - 13
SP - 8821
EP - 8824
JO - Chinese Journal of Tissue Engineering Research
JF - Chinese Journal of Tissue Engineering Research
IS - 45
ER -