Biomedical papers, 2004 (vol. 148), issue 2

Biomed. Papers 148(2), 217-220 (2004) | 10.5507/bp.2004.043

NEURAL STEM CELLS TRANSPLANTED INTO INTACT BRAINS AS NEUROSPHERES FORM SOLID GRAFTS COMPOSED OF NEURONS, ASTROCYTES AND OLIGODENDROCYTE PRECURSORS

Jana Karbanová, Jaroslav Mokrý, Lenka Kotingová
Department of Histology and Embryology, Charles University in Prague, Faculty of Medicine in Hradec Králové, Šimkova 870, P. O. Box 38, 500 38 Hradec Králové, Czech Republic

Neural stem cells (NSCs) are tissue-specific stem cells with self-renewal potential that can give rise to neurons and glia in vivo and in vitro. The aim of this study was to transplant NSCs as whole neurospheres into intact brain and assess the fate and phenotype of their progeny generated in vivo. We isolated NSCs from E14 foetal rat forebrains and cultured them in basic fibroblast and epidermal growth factor-supplemented serum-free medium in the form of neurospheres in vitro. Neurospheres were transplanted into the intact brains of 2 Wistar rats and after a period of 3 weeks, grafted brains were examined immunohistochemically. Neurospheres formed solid grafts that were found in the lateral ventricle and in the velum interpositum under the hippocampus. The majority of cells in the transplanted tissue were identified as β-III-tubulin+, NeuN+, PanNF+ and synaptophysin+ neurons and were accumulated throughout the graft centre. GFAP+ astrocytes were scattered throughout the entire graft and astrocyte processes delimited the outer and perivascular surfaces. A great number of NG2+ oligodendrocyte precursors was detected. Nestin+ endothelial cells were found to line capillaries growing in the transplant. These data indicate that nestin+ NSCs prevailing in neurospheres differentiate following transplantation into nestin- neuronal and glial cells which confirms the multipotency of NSCs. Three weeks posttranpslantation neuronal and astrocyte cells reached terminal differentiation (formation of synaptic vesicles and superficial and perivascular limiting membranes) while elements of oligodendroglial cell lineage remained immature. Grafting stem cells as non-dissociated neurospheres provide cells with favourable conditions which facilitate cell survival, proliferation and differentiation. However, in the intact brain, grafted neurosphere cells were not found to integrate with the brain parenchyma and formed a compact structure demarcated from its surroundings.

Keywords: Neural stem cells, Neurospheres, Rat, Transplantation, Differentiation, Nestin

Received: September 20, 2004; Published: December 1, 2004


References

  1. Reynolds BA, Tetzlaff W, Weiss S. (1992) A multipotent EGF responsive striatal embryonic progenitor cell produces neurons and astrocytes. J Neurosci 12, 4565-74. Go to PubMed...
  2. Davis AA, Temple S. (1994) A self-renewing multipotential stem cell in embryonic rat cerebral cortex. Nature 372, 263-6. Go to original source... Go to PubMed...
  3. Reynolds BA, Weiss S. (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255, 1707-10. Go to original source... Go to PubMed...
  4. Lendahl U, Zimmermann LB, McKay RDG. (1990) CNS stem cells express a new class of intermediate filament protein. Cell 60, 585-95. Go to original source... Go to PubMed...
  5. Vescovi AL, Reynolds BA, Fraser DD, Weiss S. (1993) bFGF regulates the proliferative fate of unipotent (neuronal) and bipotent (neuronal/astroglial) EGF-generated CNS progenitor cells. Neuron 11, 951-66. Go to original source... Go to PubMed...
  6. Gritti A, Parati EA, Cova L, Frölichsthal P, Galli R, Wanke E, Faravelli L. (1996) Multipotent stem cells from the adult mouse brain proliferate and self-renew in response to basic fibroblast growth factor. J Neurosci 16, 1091-100. Go to PubMed...
  7. Mokrý J, Němeček S. (1999) Cerebral angiogenesis shows nestin expression in endothelial cells. Gen Physiol Biophys 18 (Suppl. 1), 25-9. Go to PubMed...
  8. Wennersten A, Meijer X, Holmin S, Wahlberg L, Mathiesen T. (2004) Proliferation, migration, and differentiation of human neural stem/progenitor cells after transplantation into a rat model of traumatic brain injury. J Neurosurg 100, 88-96. Go to original source... Go to PubMed...
  9. Svendsen CN, ter Borg MG, Armstrong RJE, Rosser AE, Chandran S, Ostenfeld T, Caldwell CA. (1998) A new method for the rapid and long term growth of human neural precursor cells. J Neurosci Methods 85, 141-52. Go to original source... Go to PubMed...
  10. Burnstain, RM, Foltynie T, He X, Menon, DK, Svendsen CN, Caldwell MA. (2004) Differentiation and migration of long term expanded human neural progenitors in a partial lesion model of Parkinsons disease. Int J Biochem Cell Biol 36, 702-13. Go to original source... Go to PubMed...