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中国生物工程杂志

China Biotechnology
China Biotechnology  2009, Vol. 29 Issue (04): 39-45    DOI:
    
Impact of serially low density passage on Oct-4 positive percentage and neurogenesis potential of mouse ES derived progenitors in differentiated expanding stage
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Abstract  

Objective: To identify whether serially low density passage could decrease the Oct-4 positive cell percentage and maintain the ability of neurogenesis of mouse ES derived progenitors in expanding stage. Methods: Mouse ES were differentiated into neurons according to fivestep method and given more than 10 continuous passage by low seeding density in proliferation stage. And then cell immunohistochemistry identified Oct-4 positive cells, neurons and astrocytes, flow cytometry tested the Oct-4 positive cell percentage, and the apoptosis testing kit tested the cell apoptosis. Results: The Oct-4 positive cell percentage of mouse ES derived progenitors depressed to 4.33(1.63% from 16.17(4.8%, cell apoptosis rate of serially low and high density passage in differentiated expanding stage are respective 15.16(3.64% and 11.88(2.63%, and cell immunohistochemistry identified that GFAP and Tuj-1 of the differentiated cells are negative. Conclusion: The Oct-4 positive cell percentage of mouse ES derived progenitors decreased significantly in this differentiation system, and it’s not due to other cells apoptosis was testified. Unexpectedly, the differentiation ability of the progenitors into GFAP positive astrocytes and Tuj-1 positive neurons decreased dramatically at the same time. This result suggested that high density passage play an important part in process of the progenitors differentiated into neurocytes, and the comparision between high and low density culture supplied new platform and the development direction of the neurons differentiation mechanism from ES.



Key wordsEmbryonic stem cell;Cell culture;Neurogenesis     
Received: 01 September 2008      Published: 27 April 2009
ZTFLH:  Q254  
Cite this article:

WANG Fang- Du-Qiang-An- Shu-Wan-Wan- Tun-Di- Xu-Yan-Ling- Guan-Yun-Qian- Zhang-Yu. Impact of serially low density passage on Oct-4 positive percentage and neurogenesis potential of mouse ES derived progenitors in differentiated expanding stage. China Biotechnology, 2009, 29(04): 39-45.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2009/V29/I04/39

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