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Isolation and cultivation of neural stem cells from the embryonic rat brainand spiral cord

U Sai- Li, MA Zheng- Wen, YIN Lan, LU Pei- Hua, XU Mao- Ming

Department of Neurobiology, Shanghai Second Medical University, Shanghai 200025；Ketokin Injury Research Center，Department of Neurological Surgery，Univeisity of Louisville，511 S .Floyd Street MDR616，Louisville，KY40292，USA

Abstract

The aim of this stu勿was to establish the culture system of isolation and cultivation of the neural stem cells(NSCs) from the embryonic rat brain and spinal cord .The methods of microscopic dissection，cell cul-
ture and immunofluorescence cytochemistry were used.The results are as follows.(1)In the presence of fibroblast growth factor 2(FGY- 2) and epidermal growth factor(EGF)，both brain and spinal cord-derived stem cells proliferated and expanded in vitro for 8一10 passages(over 60 d).The period of expansion resulted in a 106fold increase in brain- derived NSCs and 105- fold increase in spinal cord-derived NSCs.These proliferating cells expressed nestin.(2) In the medium containing 1%FBS，the two NSCs populations could be induced to differ entiate into neurons，astrocytes and oligodentrocytes .The percentage of neurons(f}-tubulin IIfir) differentiated from brain- derived NSCs decreased rapidly from 11 .95士2.5 % at passage 2(PZ) to 1 .97士1 .16%atpassage 5
(P5).Significant difference was shown between PZ and PS( P<0.01).The percentage of oligodentrocytes(Ripit) differentiated from brain- derived NSCs remained mostly unchanged from 8.66士2. 93 % at P2 to 9.12士1 .13%at PS .The same differentiation patterns were found in spinal cord derived NSCs.All these results indicate that both embryonic rat brain-and spinal cord derived NSCs can expand and proliferate in vitro through multiple passages，and retain the capacity to differentiate into all three major types of cells in the central nervous systom.

Key words: central neural system;neural stem cell;embryo;fibroNast growth factor -2;epidermal growth factor

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Citing This Article：

U Sai- Li, MA Zheng- Wen, YIN Lan, LU Pei- Hua, XU Mao- Ming. Isolation and cultivation of neural stem cells from the embryonic rat brainand spiral cord . Acta Physiol Sin 2003; 55 (3): (in Chinese with English abstract).