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In vitro neurogenesis by progenitor cells isolated from the adult human hippocampus

Nature Medicine volume 6, pages 271–277 (2000)Cite this article

Abstract

Neurogenesis persists in the adult mammalian hippocampus. To identify and isolate neuronal progenitor cells of the adult human hippocampus, we transfected ventricular zone-free dissociates of surgically-excised dentate gyrus with DNA encoding humanized green fluorescent protein (hGFP), placed under the control of either the nestin enhancer (E/nestin) or the Tα1 tubulin promoter (P/Tα1), two regulatory regions that direct transcription in neural progenitor cells. The resultant P/Tα1:hGFP+ and E/nestin:enhanced (E)GFP+ cells expressed βIII-tubulin or microtubule-associated protein-2; many incorporated bromodeoxyuridine, indicating their genesis in vitro. Using fluorescence-activated cell sorting, the E/nestin:EGFP+ and P/Tα1:hGFP+ cells were isolated to near purity, and matured antigenically and physiologically as neurons. Thus, the adult human hippocampus contains mitotically competent neuronal progenitors that can be selectively extracted. The isolation of these cells may provide a cellular substrate for re-populating the damaged or degenerated adult hippocampus.

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Figure 1: Adult human hippocampus contains mitotic neuronal progenitor cells.
Figure 2: The Tα1 tubulin promoter is transcriptionally active in adult hippocampal neuronal precursor cells.
Figure 3: Adult hippocampal progenitors sorted by either E/nestin:EGFP or P/Tα1:GFP give rise to neurons.
Figure 4: The nestin enhancer drives expression in nestin protein-positive adult hippocampal cells that divide and give rise to βIII-tubulin+ neurons.
Figure 5: Neural progenitors identified by P/Tα1:hGFP and E/nestin:EGFP can be isolated by FACS.
Figure 6: FACS based on P/Tα1:hGFP and E/nestin:EGFP each enrich neuronal progenitor cells from the adult hippocampus.
Figure 7: Hippocampal cells sorted by P/Tα1:hGFP develop into physiologically mature neurons.

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Acknowledgements

We thank H.M. Keyoung for comments on the manuscript. This work was supported by the National Multiple Sclerosis Society, the Aitken Charitable Trust, the G. Harold and Leila Y. Mathers Charitable Foundation, the Human Frontiers Scientific Program, and National Institute of Neurological Disorders and Stroke grants R01NS29813 and R01NS33106.

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Authors and Affiliations

  1. Departments of Neurology and Neuroscience, Cornell University Medical College, 1300 York Ave. Room E607, New York, 10021, New York, USA

    Neeta Singh Roy, Su Wang, Abdellatif Benraiss, Catherine Harrison-Restelli & Steven A. Goldman

  2. Department of Neurosurgery, Cornell University Medical College, 1300 York Ave. Room E607, New York, 10021, New York, USA

    Richard A. R. Fraser

  3. Department of Neurosurgery, Valhalla, 10585, New York, USA

    William T. Couldwell & Maiken Nedergaard

  4. Department of Cell Biology, New York Medical College, Valhalla, 10585, New York, USA

    Li Jiang, Jian Kang & Maiken Nedergaard

  5. Division of Neuroanatomy, Osaka University Graduate School of Medicine, 2-2 Yamadaoka Suita, Osaka , 565-0867, Japan

    Ayano Kawaguchi & Hideyuki Okano

  6. Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, 2-2 Yamadaoka Suita , Osaka, 565-0867, Japan

    Ayano Kawaguchi & Hideyuki Okano

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Correspondence to Steven A. Goldman.

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Roy, N., Wang, S., Jiang, L. et al. In vitro neurogenesis by progenitor cells isolated from the adult human hippocampus. Nat Med 6, 271–277 (2000). https://doi.org/10.1038/73119

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