Culture of Mouse Neural Stem Cell Precursors

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This video describes the method used for isolation of neuroprecursors from the developing cortex of embryonic mice. The procedure for removing embryos from the uterus, dissecting the cortical tissue, and digesting the isolated cerebral cortex is shown.

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Currle, D. S., Hu, J. S., Kolski-Andreaco, A., Monuki, E. S. Culture of Mouse Neural Stem Cell Precursors. J. Vis. Exp. (2), e152, doi:10.3791/152 (2007).


Primary neural stem cell cultures are useful for studying the mechanisms underlying central nervous system development. Stem cell research will increase our understanding of the nervous system and may allow us to develop treatments for currently incurable brain diseases and injuries. In addition, stem cells should be used for stem cell research aimed at the detailed study of mechanisms of neural differentiation and transdifferentiation and the genetic and environmental signals that direct the specialization of the cells into particular cell types. This video demonstrates a technique used to disaggregate cells from the embryonic day 12.5 mouse dorsal forebrain. The dissection procedure includes harvesting E12.5 mouse embryos from the uterus, removing the "skin" with fine dissecting forceps and finally isolating pieces of cerebral cortex. Following the dissection, the tissue is digested and mechanically dissociated. The resuspended dissociated cells are then cultured in "stem cell" media that favors growth of neural stem cells.


  1. Mouse neural precursors (NPCs) were isolated from E12.5 embryo cortex.
  2. Skin and mesenchymal layers were removed from dissected telencephalic vesicles.
  3. Vesicles were incubated in 0.05% trypsin with 0.02% EDTA and 0.2% BSA in HBSS for 20 minutes at 37°C.
  4. Trypsinization was stopped by an equal volume of 1 mg/ml soybean trypsin inhibitor (Sigma #T6522) in HBSS.
  5. Tissue digests were dissociated using several rounds of trituration with fire-polished Pasteur pipettes.
  6. Cells were washed once with 0.2% BSA in HBSS and plated at 50,000 cells/ml on laminin-coated coverslips in media with 20 ng/ml EGF, 10 ng/ml FGF2 (R&D Systems or Peprotech), and 2 ug/ml heparin (Sigma).

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Great advances in our understanding of CNS development and stem cell biology have been made possible by our ability to harvest, isolate and culture embryonic neural stem cells. This video demonstrates the dissection of E12.5 mouse cerebral cortex and the subsequent disaggregation and culturing of embryonic neural stem cells. Many other other similar methods have been successfully employed by other investigators.

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Name Type Company Catalog Number Comments
Straight Iris Scissors Tool Fine Science Tools 14060-11
Medium Scissors Tool Fine Science Tools 15024-10
Micro Scissors Tool Fine Science Tools 15002-08
Bent Forceps Tool Fine Science Tools 11251-35



  1. Currle, D., Cheng, X., Hsu, C., Monuki, E. Direct and indirect roles of CNS dorsal midline cells in choroid plexus epithelial formation. Development. 132, (15), 3549-3559 (2005).
  2. Flanagan, L., Rebaza, L., Derzic, S., Schwartz, P., Monuki, E. Regulation of human neural precursor cells by laminin and integrins. J Neurosci Res. 83, 845-856 (2006).



  1. There are lots of mistakes about working in a steril conditions, the cap of the bottles should be plased upsode down.  No good lab practoce !

    Posted by: Maryam M.
    April 20, 2009 - 2:31 PM
  2. Thanks, but no contamination in 6 years.

    Posted by: Anonymous
    April 20, 2009 - 4:34 PM
  3. I think it would be better to say "Culture of Mouse neural Stem (precursor) cells". You can not use both stem and precursor terms together.

    Posted by: Hassan A.
    July 13, 2010 - 9:25 PM

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