title: High efficacy of clonal growth and expansion of adult neural stem cells.
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.
authors.affiliation: Volkswagen-Foundation Junior Group, University of Regensburg, Germany.
authors.forename: Frank-Peter
authors.initials: FP
authors.lastname: Wachs
authors.forename: Sebastien
authors.initials: S
authors.lastname: Couillard-Despres
authors.forename: Maren
authors.initials: M
authors.lastname: Engelhardt
authors.forename: Daniel
authors.initials: D
authors.lastname: Wilhelm
authors.forename: Sonja
authors.initials: S
authors.lastname: Ploetz
authors.forename: Maurice
authors.initials: M
authors.lastname: Vroemen
authors.forename: Johanna
authors.initials: J
authors.lastname: Kaesbauer
authors.forename: Goekhan
authors.initials: G
authors.lastname: Uyanik
authors.forename: Jochen
authors.initials: J
authors.lastname: Klucken
authors.forename: Claudia
authors.initials: C
authors.lastname: Karl
authors.forename: Johanna
authors.initials: J
authors.lastname: Tebbing
authors.forename: Clive
authors.initials: C
authors.lastname: Svendsen
authors.forename: Norbert
authors.initials: N
authors.lastname: Weidner
authors.forename: Hans-Georg
authors.initials: HG
authors.lastname: Kuhn
authors.forename: Juergen
authors.initials: J
authors.lastname: Winkler
authors.forename: Ludwig
authors.initials: L
authors.lastname: Aigner
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D002490:Central Nervous System / Q000166:cytology / Q000502:physiology; D002999:Clone Cells; D004247:DNA / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D007150:Immunohistochemistry; D007621:Karyotyping; D019394:Ki-67 Antigen / Q000378:metabolism; D009474:Neurons / Q000502:physiology* / Q000637:transplantation; D051381:Rats; D011916:Rats, Inbred F344; D013194:Staining and Labeling; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000502:physiology*
chemical_list: D019394:Ki-67 Antigen; D004247:DNA