Transfection of primary embryonic chicken retinal cells using cationic lipid.
Primary neuronal culture and transfection are useful tools in determining gene functions within specific tissue contexts and developmental stages. Chicken embryonic retinal cultures are easily obtainable and often robust as the chicken eye is relatively large compared to mouse eye at similar developmental stages. Various DNA-based constructs have been developed to overexpress or knockdown genes of interest and can be delivered into the cells using lipofectamine, a cationic lipid-based transfection system. Here, we describe a method to culture and transfect primary chicken embryonic retinal cells in order to manipulate genes involved in retinal development. This technique can simultaneously deliver multiple genes without construct-size constrains and permit the usage of tissue or cell type-specific promoters, and is thus a useful approach to explore gene functions during neural retina differentiation.
Department of Developmental Biology, Cincinnati Children's Research Foundation, Cincinnati, OH, USA.
Yi-Wen
YW
Hsieh
Xian-Jie
XJ
Yang
D000818:Animals; D002642:Chick Embryo; D004247:DNA / Q000378:metabolism; D007150:Immunohistochemistry; D008055:Lipids; D012160:Retina / Q000166:cytology / Q000378:metabolism*; D046509:Tissue Culture Techniques; D014162:Transfection
D008055:Lipids; C086724:Lipofectamine; D004247:DNA