Neural stem-like cell line derived from a nonhematopoietic population of human umbilical cord blood.

The ability of stem and progenitor cells to proliferate and differentiate into other lineages is widely viewed as a characteristic of stem cells. Previously, we have reported that cells from a CD34(-) (nonhematopoietic) adherent subpopulation of human cord blood can acquire a feature of multipotential neural progenitors in vitro. In the present study, using these cord blood-derived stem cells, we have established a clonal cell line termed HUCB-NSCs (human umbilical cord blood-neural stem cells) that expresses several neural antigens and has been grown in culture for more than 60 passages. During this time, HUCB-NSCs retained their growth rate, the ability to differentiate into neuronal-, astrocyte-, and oligodendrocyte-like cells and displayed a stable karyotype. DNA microarray analysis of HUCB-NSCs revealed enhanced expression of selected genes encoding putative stem and progenitor cell markers when compared to other mononuclear cells. dBcAMP-induced HUCBNSCs were further differentiated into more advanced neuronal cells. This is the first report of the establishment and characterization of a nontransformed HUCB-NSC line that can be grown continuously in a monolayer culture and induced to terminal differentiation. These cells should further our understanding of the regulatory mechanisms involved in NSC self-renewal and differentiation.

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

[2]  G. Comi,et al.  Neuronal generation from somatic stem cells: current knowledge and perspectives on the treatment of acquired and degenerative central nervous system disorders. , 2003, Current gene therapy.

[3]  M. Lucero,et al.  Immunocytochemical and physiological characterization of a population of cultured human neural precursors. , 2000, Journal of neurophysiology.

[4]  Catherine M. Verfaillie,et al.  Pluripotency of mesenchymal stem cells derived from adult marrow , 2002, Nature.

[5]  L. Denner,et al.  Production of stem cells with embryonic characteristics from human umbilical cord blood , 2005, Cell proliferation.

[6]  K. Takiguchi-Hayashi.  In vitro clonal analysis of rat cerebral cortical neurons expressing latexin, a subtype-specific molecular marker of glutamatergic neurons. , 2001, Brain research. Developmental brain research.

[7]  Clive N Svendsen,et al.  A new method for the rapid and long term growth of human neural precursor cells , 1998, Journal of Neuroscience Methods.

[8]  N. Forraz,et al.  Umbilical cord blood stem cells can expand hematopoietic and neuroglial progenitors in vitro. , 2004, Experimental cell research.

[9]  H. Varmus,et al.  Nuclear-cytoplasmic shuttling of Axin regulates subcellular localization of beta-catenin. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Wolfger von der Behrens,et al.  Transient calretinin expression defines early postmitotic step of neuronal differentiation in adult hippocampal neurogenesis of mice , 2003, Molecular and Cellular Neuroscience.

[11]  L. Espinosa,et al.  Phosphorylation by Glycogen Synthase Kinase-3β Down-regulates Notch Activity, a Link for Notch and Wnt Pathways* , 2003, Journal of Biological Chemistry.

[12]  P. Maher,et al.  High affinity immunoreactive FGF receptors in the extracellular matrix of vascular endothelial cells--implications for the modulation of FGF-2 , 1995, The Journal of cell biology.

[13]  N. Perrimon,et al.  The Promise and Perils of Wnt Signaling Through β-Catenin , 2002, Science.

[14]  R. Poulsom,et al.  Adult stem cell plasticity , 2002, The Journal of pathology.

[15]  A. Björklund,et al.  Generation and Transplantation of EGF-Responsive Neural Stem Cells Derived from GFAP–hNGF Transgenic Mice , 1997, Experimental Neurology.

[16]  S. Weiss,et al.  The Ciliary Neurotrophic Factor/Leukemia Inhibitory Factor/gp130 Receptor Complex Operates in the Maintenance of Mammalian Forebrain Neural Stem Cells , 2001, The Journal of Neuroscience.

[17]  P. Portin.  General outlines of the molecular genetics of the Notch signalling pathway in Drosophila melanogaster: a review. , 2002, Hereditas.

[18]  R. McKay,et al.  Ascorbic acid increases the yield of dopaminergic neurons derived from basic fibroblast growth factor expanded mesencephalic precursors , 2001, Journal of neurochemistry.

[19]  F. Gage,et al.  Genetic and functional differences between multipotent neural and pluripotent embryonic stem cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Y. Ha,et al.  Neural phenotype expression of cultured human cord blood cells in vitro , 2001, Neuroreport.

[21]  E. Parati,et al.  Isolation and Cloning of Multipotential Stem Cells from the Embryonic Human CNS and Establishment of Transplantable Human Neural Stem Cell Lines by Epigenetic Stimulation , 1999, Experimental Neurology.

[22]  R. Berezney,et al.  Integrative nuclear FGFR1 signaling (INFS) as a part of a universal “feed‐forward‐and‐gate” signaling module that controls cell growth and differentiation , 2003, Journal of cellular biochemistry.

[23]  D. Hess,et al.  Hematopoietic origin of microglial and perivascular cells in brain , 2004, Experimental Neurology.

[24]  Austin G Smith,et al.  Niche-Independent Symmetrical Self-Renewal of a Mammalian Tissue Stem Cell , 2005, PLoS biology.

[25]  A. Hampl,et al.  Expression and Potential Role of Fibroblast Growth Factor 2 and Its Receptors in Human Embryonic Stem Cells , 2005, Stem cells.

[26]  P. Greengard,et al.  Maintenance of pluripotency in human and mouse embryonic stem cells through activation of Wnt signaling by a pharmacological GSK-3-specific inhibitor , 2004, Nature Medicine.

[27]  Jiang Li,et al.  Gene expression in human neural stem cells: effects of leukemia inhibitory factor , 2003, Journal of neurochemistry.

[28]  John K. Heath,et al.  Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides , 1988, Nature.

[29]  D. Metcalf.  The Unsolved Enigmas of Leukemia Inhibitory Factor , 2003, Stem cells.

[30]  W. Wilkison,et al.  Neurogenic differentiation of murine and human adipose-derived stromal cells. , 2002, Biochemical and biophysical research communications.

[31]  A. Sadikot,et al.  Neurogenesis and stereological morphometry of calretinin‐immunoreactive GABAergic interneurons of the neostriatum , 2004, The Journal of comparative neurology.

[32]  S. Mahata,et al.  Neuroendocrine Cell Type‐Specific and Inducible Expression of Chromogranin/Secretogranin Genes , 2002, Annals of the New York Academy of Sciences.

[33]  B. Crain,et al.  Transplanted bone marrow generates new neurons in human brains , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[34]  M. Rao,et al.  Transdifferentiation—fact or artifact , 2003, Journal of cellular biochemistry.

[35]  J. Sanchez-Ramos,et al.  Brain as the Sea of Marrow , 2003, Experimental Neurology.

[36]  B. Pieńkowska-Grela,et al.  Overrepresentation of the short arm of chromosome 12 in seminoma and nonseminoma groups of testicular germ cell tumors. , 2002, Cancer genetics and cytogenetics.

[37]  D J Prockop,et al.  Marrow stromal cells migrate throughout forebrain and cerebellum, and they differentiate into astrocytes after injection into neonatal mouse brains. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[38]  T. Kaneko,et al.  Organization and development of corticocortical associative neurons expressing the orphan nuclear receptor Nurr1 , 2003, The Journal of comparative neurology.

[39]  J. Thomson,et al.  Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells , 2005, Nature Methods.

[40]  R. Salvi,et al.  Voltage‐Sensitive and Ligand‐Gated Channels in Differentiating Neural Stem–Like Cells Derived from the Nonhematopoietic Fraction of Human Umbilical Cord Blood , 2005, Stem cells.

[41]  F. Gage,et al.  Epigenetic control of neural stem cell fate. , 2004, Current opinion in genetics & development.

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

[43]  A. Sarnowska,et al.  Neurogenic potential of human umbilical cord blood: Neural‐like stem cells depend on previous long‐term culture conditions , 2006, Journal of neuroscience research.

[44]  C. Bever,et al.  Hematopoietic progenitors express neural genes , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[45]  M. Rao.  Conserved and divergent paths that regulate self-renewal in mouse and human embryonic stem cells. , 2004, Developmental biology.

[46]  R. Barker,et al.  Migration and differentiation of transplanted human neural precursor cells , 2003, Neuroreport.

[47]  Michael Chopp,et al.  Expression of Neural Markers in Human Umbilical Cord Blood , 2001, Experimental Neurology.

[48]  X. Fang,et al.  cAMP‐induced differentiation of human neuronal progenitor cells is mediated by nuclear fibroblast growth factor receptor‐1 (FGFR1) , 2003, Journal of neurochemistry.

[49]  C. Svendsen,et al.  Neural stem cells in the developing central nervous system: implications for cell therapy through transplantation. , 2000, Progress in brain research.

[50]  Douglas A. Melton.  “Stemness”: Definitions, Criteria, and Standards , 2004 .

[51]  A. Trounson,et al.  Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro , 2000, Nature Biotechnology.

[52]  M. Lévesque,et al.  Fetal and adult human CNS stem cells have similar molecular characteristics and developmental potential. , 2000, Brain research. Molecular brain research.

[53]  P. Sonego,et al.  Neurogenic potential of human mesenchymal stem cells revisited: analysis by immunostaining, time-lapse video and microarray , 2005, Journal of Cell Science.

[54]  L. Lagneaux,et al.  Bone marrow-derived mesenchymal stem cells already express specific neural proteins before any differentiation. , 2004, Differentiation; research in biological diversity.

[55]  A. Sadikot,et al.  Isolation of multipotent adult stem cells from the dermis of mammalian skin , 2001, Nature Cell Biology.

[56]  P. Sanberg,et al.  Human Umbilical Cord Blood Cells Express Neural Antigens after Transplantation into the Developing Rat Brain , 2002, Cell transplantation.

[57]  M. Ratajczak,et al.  Stem cell plasticity revisited: CXCR4-positive cells expressing mRNA for early muscle, liver and neural cells ‘hide out’ in the bone marrow , 2004, Leukemia.

[58]  H. Lerche,et al.  Efficient generation of neural stem cell-like cells from adult human bone marrow stromal cells , 2004, Journal of Cell Science.

[59]  M. Blasco,et al.  Long-term molecular and cellular stability of human neural stem cell lines. , 2004, Experimental cell research.

[60]  W. M. Roberts,et al.  Calretinin modifies presynaptic calcium signaling in frog saccular hair cells , 2000, Nature Neuroscience.

[61]  P. Wernet,et al.  A New Human Somatic Stem Cell from Placental Cord Blood with Intrinsic Pluripotent Differentiation Potential , 2004, The Journal of experimental medicine.

[62]  Scott Pollack,et al.  Growth factors regulate the survival and fate of cells derived from human neurospheres , 2001, Nature Biotechnology.

[63]  L. Buzanska,et al.  Human cord blood-derived cells attain neuronal and glial features in vitro. , 2002, Journal of cell science.

[64]  J. Inazawa,et al.  Damaged epithelia regenerated by bone marrow–derived cells in the human gastrointestinal tract , 2002, Nature Medicine.

[65]  A. Ruifrok,et al.  Hepatocytes and epithelial cells of donor origin in recipients of peripheral-blood stem cells. , 2002, The New England journal of medicine.