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High efficacy of clonal growth and expansion of adult neural stem cells.

Journal article
Authors Frank-Peter Wachs
Sebastien Couillard-Despres
Maren Engelhardt
Daniel Wilhelm
Sonja Ploetz
Maurice Vroemen
Johanna Kaesbauer
Goekhan Uyanik
Jochen Klucken
Claudia Karl
Johanna Tebbing
Clive Svendsen
Norbert Weidner
Hans-Georg Kuhn
Juergen Winkler
Ludwig Aigner
Published in Laboratory investigation; a journal of technical methods and pathology
Volume 83
Issue 7
Pages 949-62
ISSN 0023-6837
Publication year 2003
Published at
Pages 949-62
Language en
Keywords Animals, Cell Culture Techniques, methods, Cell Differentiation, Cell Division, Cells, Cultured, Central Nervous System, cytology, physiology, Clone Cells, DNA, analysis, Female, Flow Cytometry, Immunohistochemistry, Karyotyping, Ki-67 Antigen, metabolism, Neurons, physiology, transplantation, Rats, Rats, Inbred F344, Staining and Labeling, Stem Cell Transplantation, Stem Cells, physiology
Subject categories Cell Biology


Neural stem cells (NSCs) from the adult central nervous system are currently being investigated for their potential use in autologous cell replacement strategies. High expansion rates of NSCs in culture are crucial for the generation of a sufficient amount of cells needed for transplantation. Here, we describe efficient growth of adult NSCs in Neurobasal medium containing B27 supplement under clonal and low-density conditions in the absence of serum or conditioned medium. Expansion of up to 15-fold within 1 week was achieved on low-density NSC cultures derived from the lateral ventricle wall, the hippocampal formation, and the spinal cord of adult rats. A 27% single-cell cloning efficiency in Neurobasal/B27 combination further demonstrates its growth-promoting ability. Multipotency and nontumorgenicity of NSCs were retained despite the high rate of culture expansion. In addition, increased cell survival was obtained when Accutase, instead of trypsin, was used for enzymatic dissociation of NSC cultures. This work provides an important step toward the development of standardized protocols for highly efficient in vitro expansion of NSCs from the adult central nervous system to move more closely to the clinical use of NSCs.

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