Immobilization of erythropoietin to culture erythropoietin-dependent human leukemia cell line.
To investigate the effect of immobilized cytokine, erythropoietin (Epo) was immobilized on a culture plate and the Epo-dependent human leukemia cell line UT-7/Epo then was cultured upon the plate. A photo-reactive gelatin was mixed with Epo and the mixture was cast on a plate. The plate was then irradiated with ultraviolet light in the presence or absence of a photo-mask. After washing with water, a micropatterned or unpatterned surface was formed. A leukemia cell line dependent on Epo, UT-7/Epo, was cultured on the sample plate. On the micropatterned surface, apoptosis of cells was induced on the surface without Epo, but was not observed on the Epo-immobilized surface. This result demonstrated that Epo stimulated the cells even after immobilization. Although the activity of immobilized Epo was low, the activity was slightly higher than that achieved by soluble Epo at higher concentration. In addition, the immobilized Epo could be repeatedly used for culture of UT-7/Epo cell. The present study provided a convenient immobilization method and indicated that immobilization of cytokines will be useful for creating an artificial cell culture device.
Ito
Yoshihiro
Y
Kanagawa Academy of Science and Technology, Regenerative Medical Bioreactor Project, 3-2-1 Sakado, Takatsu-ku, Kawasaki 213-0012, Japan. y-ito@ksp.or.jp
Hasuda
Hirokazu
H
Yamauchi
Tetsuya
T
Komatsu
Norio
N
Ikebuchi
Kenji
K
D000327:Adsorption; D001672:Biocompatible Materials / Q000138:chemical synthesis / Q000528:radiation effects; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D045744:Cell Line, Tumor; D004921:Erythropoietin / Q000737:chemistry* / Q000378:metabolism*; D005780:Gelatin / Q000737:chemistry* / Q000528:radiation effects; D006801:Humans; D007938:Leukemia / Q000473:pathology* / Q000503:physiopathology*; D008422:Materials Testing; D010777:Photochemistry / Q000379:methods; D011485:Protein Binding; D023822:Tissue Engineering / Q000379:methods*; D014466:Ultraviolet Rays
D001672:Biocompatible Materials; D004921:Erythropoietin; D005780:Gelatin