Derivation of human skin fibroblast lines for feeder cells of human embryonic stem cells.
After the first derivations of human embryonic stem cell (hESC) lines on fetal mouse feeder cell layers, the idea of using human cells instead of mouse cells as feeder cells soon arouse. Mouse cells bear a risk of microbial contamination, and nonhuman immunogenic proteins are absorbed from the feeders to hESCs. Human skin fibroblasts can be effectively used as feeder cells for hESCs. The same primary cell line, which can be safely used for up to 15 passages after stock preparations, can be expanded and used for large numbers of hESC derivations and cultures. These cells are relatively easy to handle and maintain. No animal facilities or animal work is needed. Here, we describe the derivation, culture, and cryopreservation procedures of research grade human skin fibroblast lines. We also describe how to make feeder layers for hESC using these fibroblasts.
Unger
Christian
C
Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden.
Felldin
Ulrika
U
Nordenskjöld
Agneta
A
Dilber
M Sirac
MS
Hovatta
Outi
O
D002460:Cell Line; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000528:radiation effects; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000528:radiation effects; D005615:Freezing; D005720:Gamma Rays; D006801:Humans; D007223:Infant; D008297:Male; D016685:Mitomycin / Q000494:pharmacology; D008938:Mitosis / Q000187:drug effects / Q000528:radiation effects; D012867:Skin / Q000166:cytology* / Q000187:drug effects / Q000528:radiation effects
D016685:Mitomycin