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. CONCLUSION: The hNSCs isolated from the human embryonic tissues retained their biological features after long-term culture in vitro.
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. CONCLUSION: The hNSCs isolated from the human embryonic tissues retained their biological features after long-term culture in vitro.
Authors: J D Flax; S Aurora; C Yang; C Simonin; A M Wills; L L Billinghurst; M Jendoubi; R L Sidman; J H Wolfe; S U Kim; E Y Snyder Journal: Nat Biotechnol Date: 1998-11 Impact factor: 54.908
Authors: C N Svendsen; M A Caldwell; J Shen; M G ter Borg; A E Rosser; P Tyers; S Karmiol; S B Dunnett Journal: Exp Neurol Date: 1997-11 Impact factor: 5.330