Purification of a pluripotent neural stem cell from the adult mouse brain

The adult mammalian central nervous system (CNS) contains a population of neural stem cells (NSCs) with properties said to include the generation of non-neural progeny. However, the precise identity, location and potential of the NSC in situ remain unclear. We purified NSCs from the adult mouse brain by flow cytometry, and directly examined the cells' properties. Here we show that one type of NSC, which expresses the protein nestin but only low levels of PNA-binding and HSA proteins, is found in both ependymal and subventricular zones and accounts for about 63% of the total NSC activity. Furthermore, the selective depletion of the population of this stem cell in querkopf mutant mice (which are deficient in production of olfactory neurons) suggests that it acts as a major functional stem cell in vivo. Most freshly isolated NSCs, when co-cultured with a muscle cell line, rapidly differentiated in vitro into myocytes that contain myosin heavy chain (MyHC). This demonstrates that a predominant, functional type of stem cell exists in the periventricular region of the adult brain with the intrinsic ability to generate neural and non-neural cells.

[1]  M. Ikawa,et al.  Generating green fluorescent mice by germline transmission of green fluorescent ES cells , 1998, Mechanisms of Development.

[2]  S. Weiss,et al.  Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. , 1992, Science.

[3]  E. Parati,et al.  Multipotential stem cells from the adult mouse brain proliferate and self-renew in response to basic fibroblast growth factor , 1996, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[4]  G. Rougon,et al.  mCD24 expression in the developing mouse brain and in zones of secondary neurogenesis in the adult , 1996, Neuroscience.

[5]  Jonas Frisén,et al.  Identification of a Neural Stem Cell in the Adult Mammalian Central Nervous System , 1999, Cell.

[6]  R. McKay,et al.  CNS stem cells express a new class of intermediate filament protein , 1990, Cell.

[7]  S. Mckercher,et al.  Turning blood into brain: cells bearing neuronal antigens generated in vivo from bone marrow. , 2000, Science.

[8]  A. Salner,et al.  Differing stem cell self-renewal of lectin-separated murine bone marrow fractions. , 1982, Journal of the National Cancer Institute.

[9]  U. Lendahl,et al.  Generalized potential of adult neural stem cells. , 2000, Science.

[10]  L. Richards,et al.  De novo generation of neuronal cells from the adult mouse brain. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[11]  D. van der Kooy,et al.  Adult Mammalian Forebrain Ependymal and Subependymal Cells Demonstrate Proliferative Potential, but only Subependymal Cells Have Neural Stem Cell Characteristics , 1999, The Journal of Neuroscience.

[12]  E. Parati,et al.  Epidermal and Fibroblast Growth Factors Behave as Mitogenic Regulators for a Single Multipotent Stem Cell-Like Population from the Subventricular Region of the Adult Mouse Forebrain , 1999, The Journal of Neuroscience.

[13]  F. Gage,et al.  Mammalian neural stem cells. , 2000, Science.

[14]  R. McKay,et al.  Stem Cells in the Central Nervous System , 1997, Science.

[15]  R. Johnson,et al.  Dye efflux studies suggest that hematopoietic stem cells expressing low or undetectable levels of CD34 antigen exist in multiple species , 1997, Nature Medicine.

[16]  P. Bartlett Pluripotential hemopoietic stem cells in adult mouse brain. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[17]  C. Lois,et al.  Proliferating subventricular zone cells in the adult mammalian forebrain can differentiate into neurons and glia. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Giulio Cossu,et al.  Skeletal myogenic potential of human and mouse neural stem cells , 2000, Nature Neuroscience.

[19]  A. Beggs,et al.  Deficiency of a skeletal muscle isoform of α-actinin (α-actinin-3) in merosin-positive congenital muscular dystrophy , 1996, Neuromuscular Disorders.

[20]  P. Tam,et al.  The somitogenetic potential of cells in the primitive streak and the tail bud of the organogenesis-stage mouse embryo. , 1992, Development.

[21]  A. Vescovi,et al.  Turning brain into blood: a hematopoietic fate adopted by adult neural stem cells in vivo. , 1999, Science.

[22]  P. Gruss,et al.  Querkopf, a MYST family histone acetyltransferase, is required for normal cerebral cortex development. , 2000, Development.

[23]  G. Rougon,et al.  mCD24, a glycoprotein transiently expressed by neurons, is an inhibitor of neurite outgrowth , 1996, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[24]  Daniel A. Lim,et al.  Subventricular Zone Astrocytes Are Neural Stem Cells in the Adult Mammalian Brain , 1999, Cell.

[25]  H. Blau,et al.  From marrow to brain: expression of neuronal phenotypes in adult mice. , 2000, Science.