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Forschungsdatenbank PMU-SQQUID

High efficacy of clonal growth and expansion of adult neural stem cells.
Wachs, FP; Couillard-Despres, S; Engelhardt, M; Wilhelm, D; Ploetz, S; Vroemen, M; Kaesbauer, J; Uyanik, G; Klucken, J; Karl, C; Tebbing, J; Svendsen, C; Weidner, N; Kuhn, HG; Winkler, J; Aigner, L;
Lab Invest. 2003; 83(7):949-962
Originalarbeiten (Zeitschrift)

PMU-Autor/inn/en

Aigner Ludwig
Couillard-Després Sébastien

Abstract

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.


Useful keywords (using NLM MeSH Indexing)

Animals

Cell Culture Techniques/methods*

Cell Differentiation

Cell Division

Cells, Cultured

Central Nervous System/cytology

Central Nervous System/physiology

Clone Cells

DNA/analysis

Female

Flow Cytometry

Immunohistochemistry

Karyotyping

Ki-67 Antigen/metabolism

Neurons/physiology*

Neurons/transplantation

Rats

Rats, Inbred F344

Staining and Labeling

Stem Cell Transplantation

Stem Cells/physiology*