Cultivation and identification of neural stem cells cultivated by the methods of neurospheres and adherent monolayer cells

Abstract

Abstract:Objective:AIM To explore two methods of culturing mouse embryo neural stem cells in vitro and identification of their differentiation．Methods:The neural stem cells were derived from the cortex tissue of mouse embryo at gestation day of 14～16 with two methods: culturing of neurosphere in floating and culturing of monolayer adherent cells. To observe the development of the neural stem cell，the expression of the stem cell marker Nestin and the neural marker MAP-2 and the glial marker GFAP was examined with immunofluorescence.Results:The cultured neural stem cells proliferated．The maintenance of stem cell property was preserved with both methods as revealed by Nestin immunofluorescence. These stem cells are able to differentiate into glial (GFAP positive) and neuronal lineage (MAP-2positive).Conclusion:With the help of growth factors and serum-free technique，the mouse neural stem cells cultivated by methods of neural stem cell spheres and adherent monolayer cells，have potential of proliferation and differentiation.

Key words:cerebral cortex,neural stem cells,neurosphere,monolayer adherent cells

HAO Jun-Rong, ZHANG Li, QIN Zheng-Hong. Cultivation and identification of neural stem cells cultivated by the methods of neurospheres and adherent monolayer cells[J]. Journal of Materials Sciences & Technology, 2011, 1(1): 51-55.

References

1 Lee JM，Bae JS，Jin HK. Intracerebellar transplantation of neural stem cells into mice with neurodegeneration improves neuronal networks with functional synaptic transmission[J]. J Vet Med Sci，2010，72(8)：999-1009

2 Sharma R，McMillan CR，Niles LP. Neural stem cell transplantation and melatonin treatment in a 6-hydroxydopamine model of Parkinson's disease[J]. J Pineal Res，2007，43(3)：245-254

3 Bantubungi K，Blum D，Cuvelier L，et al. Stem cell factor and mesenchymal and neural stem cell transplantation in a rat model of Huntington's disease[J]. Mol Cell Neurosci. 2008，37(3)：454-470

4 钟德君，张德盛，宋跃明. 鼠胚神经干细胞体外培养方法的优化[J]. 中国组织工程研究与临床康复，2007，11(50)：10088-10092

5 Kim HT，Kim IS，Lee IS，et al. Human neurospheres derived from the fetal central nervous system are regionally and temporally specified but are not committed[J]. Exp Neurol，2006，199(1)：222–235

6 Naveiras O，Daley GQ. Stem cells and their niche：a matter of fate[J]. Cell Mol Life Sci，2006，63(7-8)：760–766

7 Seaberg RM，van der Kooy D. Adult rodent neurogenic regions：the ventricular subependyma contains neural stem cells，but the dentate gyrus contains restricted progenitors[J]. J Neurosci，2002，22(5)：1784–1793

8 Xing Cheng-ming，Geng Xiu-li，Liu Yu-xia. A new method to culture primary neural stem cells of embryonic rats[J]. 中国组织工程研究与临床康复，2008，12(16)：3189-3192

9 王霞，韦霄郑，唯韡，等. 大鼠胚胎脑新皮质神经干细胞的体外分离培养方法研究[J].卫生研究，2010，39(6)：674-681

10 余资江，应大君，崔晓萍，等. 新生小鼠海马源性神经干细胞体外分离培养细胞群的增殖分化[J]. 中国临床康复，2006，10(41)：1-4

11 丁道芳，邢三丽，周鸣鸣，等. 大鼠胚胎神经干细胞单克隆化及单层化培养和鉴定[J]. 生物化学与生物物理进展，2009，36(1)：72-76

[1]	ZHANG Li-hang， YIN Ming.Thinking of Influences of Small Molecules on Neural Stem Cell Neurogenesis[J]. Acta Neuropharmacologica, 2017, 7(1): 1-9.
[2]	LUO Lin-ming, JI Yi-na, CHEN Nai-hong.Research Progress on the Effect of Neural Stem Cells in the Occurrence and Development of Glioma[J]. Acta Neuropharmacologica, 2015, 5(5): 28-33.
[3]	WANG Lun-zheng, XIE Wen-juan, TANG Tie-Shan.Neural Stem Cells, Adult Neurogenesis and Cell Therapy for Neurodegenerative Diseases[J]. Acta Neuropharmacologica, 2015, 5(3): 46-64.
[4]	LI Xin, HUO Yu-shu.Neural Stem Cells with It’s Pharmacology[J]. Acta Neuropharmacologica, 2012, 2(2): 48-57.