Fabrication of a Novel Cell Culture System Using DNA-Grafted Substrates and DNase.
In conventional cell culture systems, trypsin is generally used for cell harvesting. However, trypsin damages the cells due to the nonselective degradation of proteins on the cell surface. This is a critical issue for cell culture systems. Therefore, an alternative cell culture system with the lowest possible impact on cells is desired. In this paper, we have focused on DNA as a sacrificial layer and DNase as an alternate enzyme instead of trypsin. DNase ought not to result in damage to or stress on cells as it only hydrolyzes DNAs while the plasma membrane and extracellular matrices are basically composed of lipids, proteins, and glycosides. Therefore, we fabricated DNA-grafted substrates as cell culture dishes and evaluated this novel cell culture system. As a result, we were able to culture several types of mammalian cells on the DNA-grafted substrates, with the cells harvested using DNase with only little damage to the cells. This cell culture system could provide a breakthrough in cell culturing technology.
Hideyuki
H
Mitomo
Asumi
A
Eguchi
Yasunobu
Y
Suzuki
Yasutaka
Y
Matsuo
Kenichi
K
Niikura
Kohji
K
Nakazawa
Kuniharu
K
Ijiro
D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002470:Cell Survival; D004247:DNA / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism*; D061252:Feeder Cells / Q000166:cytology; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D051379:Mice; D041681:NIH 3T3 Cells; D013379:Substrate Specificity; D014867:Water / Q000737:chemistry
D005456:Fluorescent Dyes; D014867:Water; D004247:DNA; D003851:Deoxyribonucleases