Human primary mixed brain cultures: preparation, differentiation, characterization and application to neuroscience research

Mol Brain. 2014 Sep 16;7:63. doi: 10.1186/s13041-014-0063-0.


Background: Culturing primary cortical neurons is an essential neuroscience technique. However, most cultures are derived from rodent brains and standard protocols for human brain cultures are sparse. Herein, we describe preparation, maintenance and major characteristics of a primary human mixed brain culture, including neurons, obtained from legally aborted fetal brain tissue. This approach employs standard materials and techniques used in the preparation of rodent neuron cultures, with critical modifications.

Results: This culture has distinct differences from rodent cultures. Specifically, a significant numbers of cells in the human culture are derived from progenitor cells, and the yield and survival of the cells grossly depend on the presence of bFGF. In the presence of bFGF, this culture can be maintained for an extended period. Abundant productions of amyloid-β, tau and proteins make this a powerful model for Alzheimer's research. The culture also produces glia and different sub-types of neurons.

Conclusion: We provide a well-characterized methodology for human mixed brain cultures useful to test therapeutic agents under various conditions, and to carry forward mechanistic and translational studies for several brain disorders.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Blotting, Western
  • Brain / cytology*
  • Cell Culture Techniques / methods*
  • Cell Differentiation* / drug effects
  • Cell Survival
  • Cells, Cultured
  • Dopaminergic Neurons / cytology
  • Dopaminergic Neurons / drug effects
  • Embryo, Mammalian / cytology
  • Humans
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Mice
  • Neural Stem Cells / cytology
  • Neural Stem Cells / drug effects
  • Neuroglia / cytology
  • Neurosciences*
  • Nucleic Acids / metabolism
  • RNA, Small Interfering / metabolism
  • Research*
  • Serotonergic Neurons / cytology
  • Serotonergic Neurons / drug effects
  • Staining and Labeling
  • Time Factors
  • Transfection


  • Biomarkers
  • Intercellular Signaling Peptides and Proteins
  • Nucleic Acids
  • RNA, Small Interfering