Prospective Identification, Isolation by Flow Cytometry, and In Vivo Self-Renewal of Multipotent Mammalian Neural Crest Stem Cells
Multipotent and self-renewing neural stem cells have been isolated in culture, but equivalent cells have not yet been prospectively identified in neural tissue. Using cell surface markers and flow cytometry, we have isolated neural crest stem cells (NCSCs) from mammalian fetal peripheral nerve. These cells are phenotypically and functionally indistinguishable from NCSCs previously isolated by culturing embryonic neural tube explants. Moreover, in vivo BrdU labeling indicates that these stem cells self-renew in vivo. NCSCs freshly isolated from nerve tissue can be directly transplanted in vivo, where they generate both neurons and glia. These data indicate that neural stem cells persist in peripheral nerve into late gestation by undergoing self-renewal. Such persistence may explain the origins of some PNS tumors in humans.
© 1999 Cell Press. Under an Elsevier user license. Received 22 December 1998, Revised 12 February 1999, Available online 27 September 2000. We thank Gaby Mosconi for laboratory management; Lan Dinh, Hieu Phan, and Ling Wang for technical assistance; and Robert Vega for animal ordering. Thanks to Rochelle Diamond and Pat Koen of the Caltech Flow-Cytometry Facility for FACS operation and to J. J. Archelos for monoclonal anti-P_0 antibody. Thanks also to Ken Weinberg and Lora Barsky for providing access to flow cytometry at Children's Hospital, Los Angeles. S. J. M. was supported by postdoctoral fellowships from the Natural Sciences and Engineering Research Council of Canada and the American Cancer Society, California Division. D. J. A. is an Investigator of the Howard Hughes Medical Institute. This work was supported by a program project grant from the NIH (Barbara Wold, P. I.).