Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (36): 6707-6710.doi: 10.3969/j.issn.1673-8225.2010.36.013

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Establishment of two-dimensional gel electrophoresis technique for embryonic rat neural stem cells

Liu Huan1, Huang Guo-wei1, He Bing2, Yang Yang1, Zhao Lin1   

  1. 1 School of Public Health, Tianjin Medical University, Tianjin  300070, China; 2 Tianjin Key Laboratory for Biomarkers of Occupational and Environmental Hazard, Medical College of Chinese People’s Armed Police Forces, Tianjin  300162, China
  • Online:2010-09-03 Published:2010-09-03
  • Contact: Huang Guo-wei, Doctor, Professor, Doctoral supervisor, School of Public Health, Tianjin Medical University, Tianjin 300070, China guoweihuang@yahoo.com.cn
  • About author:Liu Huan★, Master, Lecturer, School of Public Health, Tianjin Medical University, Tianjin 300070, China liuhuan@tijmu.edu.cn
  • Supported by:

    the National Natural Science Foundation of China, No. 30771797*; the Science Program of Tianjin Medical University, No. 2008Ky12*

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Abstract:

BACKGROUND: Neural stem cells are the current hot topic, and two-dimensional electrophoresis is the basic technique for proteome, it is necessary to establish a stable two-dimensional gel electrophoresis technology system for neural stem cells.
OBJECTIVE: To establish a stable, reproducible two-dimensional gel electrophoresis separation method for neural stem cells through analysis of several key steps of two-dimensional electrophoresis.
METHODS: Embryonic rat neural stem cells were cultured in serum-free cell culture. Proteins were extracted from samples and lysed. After passive rehydration, samples were isolated by different isoelectric focusing (IEF) parameters. Protein maps were analyzed by PDQuest8.0. Quality of two-dimensional gel electrophoresis maps under different IEF parameters, number and reproducibility of protein spots were detected.
RESULTS AND CONCLUSION: Quality of two-dimensional gel electrophoresis diagrams differs with IEF parameters. Low-voltage step could remove the cross striations. Ideal electrophoresis could be obtained under 80 000 Vh. Proper two-dimensional gel electrophoresis methods can be constructed by optimizing IEF parameters.

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