Neuronal and glial differentiation following culture of the human embryonic cortical stem cells.
OBJECTIVE
To set up long-term in vitro culture system of the human neural stem cells (hNSC) and to study their biological properties.


METHODS
Human fetuses aged about 20 weeks following spontaneous abortion were adopted. A serum-free medium containing basic fibroblast growth factor and epidermal growth factor was used to make the hNSCs divide continuously in the culture. The growth curve of continually passaged cells was examined. The effects of long-term culture on the cell cycle, cell differentiation were analyzed. The cell cycles of these cells were analyzed using flow cytometry.


RESULTS
The cells from the human embryonic cortical tissue could be maintained and propagated in the presence of growth factors. Neurospheres were generated continually. Only one month after the primary culture, the precursors could be effectively discarded. The cells could be cultured for ten months. The cells had an exponential, consistent growth. The cell cycle analysis indicated that the hNSCs maintained remarkable proliferation. Upon differentiation, the hNSCs gave rise to mature cells. The multi-lineage potential of differentiation after different passages remained unchanged.


CONCLUSIONS
The hNSCs isolated from the human embryonic tissues retained their biological features after long-term culture in vitro.
Liang
Peng
P
Department of Neurosurgery, First Medical College of Harbin Medical University, Harbin 150001, PR China. chalp@sina.com.cn
Zhao
Shiguang
S
Kawamoto
Keiji
K
Jin
Lianhong
L
Liu
Enzhong
E
D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology* / Q000196:embryology; D003470:Culture Media; D004622:Embryo, Mammalian / Q000166:cytology*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D039902:Multipotent Stem Cells / Q000166:cytology*; D009457:Neuroglia / Q000166:cytology*; D009474:Neurons / Q000166:cytology*; D013997:Time Factors
D003470:Culture Media; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor