Effect of basic fibroblast growth factor on cultured rat neural stem cell in three-dimensional collagen gel.
OBJECTIVE
The effect of growth factors on the three-dimensional culture of neural stem cells has not been reported. We studied the effect of basic fibroblast growth factor (bFGF) on cultured rat neural stem cells in a three-dimensional culture.


METHODS
We cultured rat neural stem cells in collagen gel matrix for three-dimensional culture and examined the effect of bFGF under such culture conditions.


RESULTS
After 4 days culture, the cell density in the bFGF treatment group was 12 times that of the non-treatment group, reaching a significantly high value. In the bFGF treatment group, microtubule associate protein (MAP)-2-positive cell aggregation occurred, although in the bFGF non-treatment group there was no MAP-2-positive cell aggregation and few of the cells were sparsely distributed. Also, in the bFGF treatment group, MAP-2-positive cell aggregation had a luminal structure similar to neural rosettes. There was elongation of MAP-2-positive neurites from the cell aggregation to the circumference in the bFGF treatment group.


CONCLUSIONS
bFGF is known to induce the proliferation, but not the differentiation of neural stem cells in two-dimensional cultures. However, in the three-dimensional culture, bFGF induced both the proliferation and differentiation of neural stem cells. The three-dimensional culture is, therefore, considered a useful method for predicting the response of neural stem cells to cytokines or biologically active substances in vivo.
Itoh
Tatsuki
T
Department of Pathology, Kinki University School of Medicine, Osaka, Japan. tatsuki@med.kindai.ac.jp
Satou
Takao
T
Dote
Kensaku
K
Hashimoto
Shigeo
S
Ito
Hiroyuki
H
D000818:Animals; D002452:Cell Count / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen; D046508:Culture Techniques / Q000379:methods; D004622:Embryo, Mammalian; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005782:Gels; D007150:Immunohistochemistry / Q000379:methods; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013234:Stem Cells / Q000187:drug effects* / Q000378:metabolism
D005782:Gels; C489787:MAP2 protein, rat; D008869:Microtubule-Associated Proteins; D016222:Fibroblast Growth Factor 2; D003094:Collagen