Culture and differentiation of cynomolgus monkey neural stem cells
10.3969/j.issn.1673-8225.2009.45.003
- VernacularTitle:食蟹猴神经干细胞的培养与分化
- Author:
Sunxing YAN
;
Xiaoming LIU
;
Peng XIANG
;
Peng WANG
;
Ding WANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(45):8821-8824
- CountryChina
- Language:Chinese
-
Abstract:
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 Canter 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 cm~2 culture flask with concentration of (2.0-5.0)×10~5.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 passagad 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-omithine/fibronectin pre-coated dish,neurospheras adhered to the dish and many cells migrated out of neurnshperes and proliferated as monolayer.The immunocytochemistry showed that NSCs which culture medium contain EGF and bFGF preserve neatin positive,and the cmNSCs that without EGF and bFGF differentiated into astrocytes (GFAP positive),neurons (Tuj-1 positive) and oligodendrocytes (O4 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.
