Application of genetically modified feeder cells for culture of keratinocytes.
In living bodies, there are many cell-cell interactions, including epithelial-mesenchymal interactions, to control cell growth and differentiation, and disturbances to such systems are believed to be a critical reason for many diseases, including cancer. Recently, many growth factors mediating epithelial- mesenchymal interactions have been revealed by molecular biological research. These growth factors are critical tools for both basic research and medical applications. However, obtaining purified forms of such growth factor proteins is difficult. To conveniently analyze the biological effects of such factors on keratinocytes, in this protocol, we describe how to introduce ectopic genes of growth factors of interest (sonic hedgehog and wnt-3 genes as examples) into Swiss-3T3 cells, one of the most widely used feeder cells for epithelial cell culture, and how to coculture the keratinocytes with them.
Kameda
Takashi
T
Department of Biochemistry, Akita University, School of Medicine, Akita, Japan.
Sugiyama
Toshihiro
T
D016475:3T3 Cells; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000737:chemistry; D000078404:Epidermal Cells; D004847:Epithelial Cells / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology; D015603:Keratinocytes / Q000166:cytology*; D051379:Mice; D011506:Proteins / Q000378:metabolism; D012867:Skin; D051153:Wnt Proteins; D060508:Wnt3 Protein
D017077:Culture Media, Conditioned; D011506:Proteins; D051153:Wnt Proteins; D060508:Wnt3 Protein; C555508:Wnt3 protein, mouse