Abstract
BackgroundCulturing 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.ResultsThis 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.ConclusionWe 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s13041-014-0063-0) contains supplementary material, which is available to authorized users.
Highlights
Culturing primary cortical neurons is an essential neuroscience technique
Rodents normally do not experience human-like neurodegenerative disorders such as Alzheimer’s disease (AD) or Parkinson’s disease (PD)
We have recently developed and characterized a primary neuron culture derived from the parenchyma of legally aborted human fetal brains
Summary
Culturing primary cortical neurons is an essential neuroscience technique. most cultures are derived from rodent brains and standard protocols for human brain cultures are sparse. 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. The applicability of our culture to studying neurodegenerative disorders has been shown by our ability to demonstrate the presence of several neuronal and synaptic markers and of significant levels of Alzheimer’s Aβ peptides, and the transfectability of small RNA species, such as siRNAs. we are able to outline a protocol for a culture that displays: a) both neuronal and glial characteristics; b) physiological activity; c) the potential for extensive utility across neuroscience. We have successfully employed the primary culture system described in this protocol in our research workflow [19]
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