TY - JOUR
T1 - Selective and rapid expansion of human neural progenitor cells on substrates with terminally anchored growth factors
AU - Konagaya, Shuhei
AU - Kato, Koichi
AU - Nakaji-Hirabayashi, Tadashi
AU - Iwata, Hiroo
N1 - Funding Information:
Part of this study was supported by a Grant-in-Aid for Scientific Research ( 22300164 ) from the Ministry of Education, Culture, Sports, Science, and Technology , Japan and by the Cooperative Research Program of Institute for Frontier Medical Sciences, Kyoto University , Japan.
PY - 2013/8
Y1 - 2013/8
N2 - Human neural progenitor cells (hNPCs) are a potential source for cell transplantation therapy in central nervous disorders. Neurosphere culture, the standard method for obtaining hNPCs, suffers from several limitations including the heterogeneity of cells in a neurosphere and the limitation of growth rate due to the presence of differentiated cells in the neurospheres. To overcome these limitations, we developed culture substrates that enable the selective expansion of hNPCs in adherent culture. Epidermal growth factor and basic fibroblast growth factor were fused with hexahistidine (EGF-His and bFGF-His, respectively) and were immobilized alone or in combination onto Ni ion-bound glass through coordination. When hNPCs derived from human fetal brain were cultured on these substrates, adhesion and proliferation of hNPCs took place most efficiently on the substrate with both EGF-His and bFGF-His compared to substrates with either factor alone and to a control substrate without growth factors. The rate of cell proliferation was two-fold higher in the adherent culture on the substrate immobilized with both EGF-His and bFGF-His than in the standard neurosphere culture. A cell population obtained after 5 days of culture on the substrate contained nestin-expressing progenitors (>90%). We conclude that the culture substrate with co-immobilized EGF and bFGF is effective for the selective expansion of hNPCs.
AB - Human neural progenitor cells (hNPCs) are a potential source for cell transplantation therapy in central nervous disorders. Neurosphere culture, the standard method for obtaining hNPCs, suffers from several limitations including the heterogeneity of cells in a neurosphere and the limitation of growth rate due to the presence of differentiated cells in the neurospheres. To overcome these limitations, we developed culture substrates that enable the selective expansion of hNPCs in adherent culture. Epidermal growth factor and basic fibroblast growth factor were fused with hexahistidine (EGF-His and bFGF-His, respectively) and were immobilized alone or in combination onto Ni ion-bound glass through coordination. When hNPCs derived from human fetal brain were cultured on these substrates, adhesion and proliferation of hNPCs took place most efficiently on the substrate with both EGF-His and bFGF-His compared to substrates with either factor alone and to a control substrate without growth factors. The rate of cell proliferation was two-fold higher in the adherent culture on the substrate immobilized with both EGF-His and bFGF-His than in the standard neurosphere culture. A cell population obtained after 5 days of culture on the substrate contained nestin-expressing progenitors (>90%). We conclude that the culture substrate with co-immobilized EGF and bFGF is effective for the selective expansion of hNPCs.
KW - Central nervous system
KW - Growth factors
KW - Neural progenitor cells
KW - Regenerative medicine
KW - Surface immobilization
UR - http://www.scopus.com/inward/record.url?scp=84878589063&partnerID=8YFLogxK
U2 - 10.1016/j.biomaterials.2013.04.041
DO - 10.1016/j.biomaterials.2013.04.041
M3 - 学術論文
C2 - 23683722
AN - SCOPUS:84878589063
SN - 0142-9612
VL - 34
SP - 6008
EP - 6014
JO - Biomaterials
JF - Biomaterials
IS - 25
ER -