High efficacy of clonal growth and expansion of adult neural stem cells.
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.
Wachs
Frank-Peter
FP
Volkswagen-Foundation Junior Group, University of Regensburg, Germany.
Couillard-Despres
Sebastien
S
Engelhardt
Maren
M
Wilhelm
Daniel
D
Ploetz
Sonja
S
Vroemen
Maurice
M
Kaesbauer
Johanna
J
Uyanik
Goekhan
G
Klucken
Jochen
J
Karl
Claudia
C
Tebbing
Johanna
J
Svendsen
Clive
C
Weidner
Norbert
N
Kuhn
Hans-Georg
HG
Winkler
Juergen
J
Aigner
Ludwig
L
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*
D019394:Ki-67 Antigen; D004247:DNA