Fabrication of wool keratin sponge scaffolds for long-term cell cultivation.
Wool keratin sponge scaffolds were fabricated by lyophilization of an aqueous wool keratin solution after controlled freezing. Freezing at -20 degrees C for 3 days was needed for the preparation of stable sponges, which did not show significant changes against heat treatment at 60 degrees C for several hours. Scanning electron microscopic observation revealed that the wool keratin sponges had a homogeneously porous microstructure, the pore size was approximately equal to 100 microm. At 1 h from seeding, the adhesion of cells was observed and at 1 day, cells spread on the sponge surface. Rapid cell growth on the sponge (doubling time: 29.0 h) was observed for at least 7 days, as well as on a commercially available plastic culture dish (doubling time: 27.4 h). At long-term (23-43 days) cultivation, cells were constantly counted to be approximately 4.2-7.4 million per sponge (1 cm in diameter). The maximum cell number was 7.4 million, approximately 37 times higher than on the same area dish. Living cells on the sponge were observed at 23-43 days by SEM observation and no abnormal morphology of the cells was observed. These results show that wool keratin sponges are useful scaffolds for long-term and high-density cell cultivation.
Tachibana
Akira
A
Department of Bioapplied Chemistry, Graduate School of Engineering, Osaka City University, Sugimoto 3-3-138, Sumiyoshi-ku, Osaka 558-8585, Japan. tatibana@bioa.eng.osaka-cu.ac.jp
Furuta
Yasunari
Y
Takeshima
Hideyuki
H
Tanabe
Toshizumi
T
Yamauchi
Kiyoshi
K
D000818:Animals; D018929:Cell Culture Techniques; D002455:Cell Division; D002477:Cells / Q000166:cytology*; D007633:Keratins; D008855:Microscopy, Electron, Scanning; D014935:Wool
D007633:Keratins