Neuron-like differentiation and selective ablation of undifferentiated embryonic stem cells containing suicide gene with Oct-4 promoter.
In vivo transplantation of undifferentiated embryonic stem (ES) cells can produce teratomas with uncontrolled cell proliferation. Although ES cells may be attractive candidates for human cell-replacement therapy in the future, the major limitation of its application to the therapy is teratoma formation. In the present study, ES cells containing herpes simplex virus-thymidine kinase (HSV-tk) transgene for a suicide gene expression under the control of the Oct-4 promoter was used for ablation of undifferentiated ES cells, which may produce teratomas, using three-dimensional cell culture system allowing a multilayer cell construct. Selective ablation of undifferentiated ES cells expressing HSV-tk gene under the control of Oct-4 promoter was achieved by ganciclovir treatment. Surviving ES cells after ganciclovir treatment expressed several neuron-associated markers such as synaptophysin, beta-tubulin, vesicular glutamate transporter 1, syntaxin, protein kinase C and glial fibrillary acidic protein (GFAP) but not Oct-4. Coexpression of synaptophysin as a marker of neuronal synapse and GFAP as that of glial fibers in the surviving ES cells revealed finely structured neuronal network. Furthermore, decrease of Ki-67 proliferative index was detected in the surviving ES cells. In conclusion, selective ablation of undifferentiated ES cells by a suicide gene decreases proliferative activity and induces neuron-like differentiation in ES cells.
Hara
Akira
A
Department of Tumor Pathology, Gifu University Graduate School of Medicine, Gifu, Japan. ahara@gifu-u.ac.jp
Aoki
Hitomi
H
Taguchi
Ayako
A
Niwa
Masayuki
M
Yamada
Yasuhiro
Y
Kunisada
Takahiro
T
Mori
Hideki
H
D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000235:genetics; D000998:Antiviral Agents / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000201:enzymology*; D015774:Ganciclovir / Q000494:pharmacology; D041722:Genes, Transgenic, Suicide / Q000235:genetics; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000201:enzymology*; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D018139:Simplexvirus / Q000201:enzymology*; D013937:Thymidine Kinase / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics
D000943:Antigens, Differentiation; D000998:Antiviral Agents; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D014764:Viral Proteins; D013937:Thymidine Kinase; D015774:Ganciclovir