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(American Journal of Pathology. 2002;161:1567-1576.)
© 2002 American Society for Investigative Pathology

Technical Advance

Isolation of Living Neurons from Human Elderly Brains Using the Immunomagnetic Sorting DNA-Linker System

Yoshihiro Konishi*, Kristina Lindholm*, Li-Bang Yang*, Rena Li*{dagger} and Yong Shen*{ddagger}

From the Haldeman Laboratory of Molecular and Cellular Neurobiology* and the Roberts Center for Alzheimer’s Research,{dagger}0 Sun Health Research Institute, Sun City; and the Molecular and Cellular Biology Program,{ddagger} Arizona State University, Tempe, Arizona

Isolation and culture of mature neurons from affected brain regions during diseased states provide a well-suited in vitro model system to study age-related neurodegeneration under dynamic conditions at cellular levels. We have developed a novel technique to isolate living neurons from rapidly autopsied human elderly brains, and have succeeded in keeping them alive in vitro. Specifically, the parietal cortex blocks were fractionated by density gradients and further enriched for neurons by an immunomagnetic sorting DNA-linker technique. The postmortem interval averaged 2.6 hours. After isolation and purification of neurons using this technology, the cells were maintained in vitro for 2 weeks. Our evaluation revealed that 80% of the isolated cells were neurons and they exhibited neurotransmitter phenotypes (glutamate and {gamma}-aminobutyric acid) as well as glutamate receptors. Studies on cell viability and calcium influx suggest that these isolated living cortical neurons still retain their typical neuronal functions. Our present study demonstrates that neurons isolated from human elderly brain autopsies can survive in vitro and maintain their functional properties. Our study has opened an opportunity to apply such neurons to dynamic pharmacological studies of neurological disorders at the single-cell level.




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