Generation of induced pluripotent stem cells from mouse cancer cells.
Reprogramming of cancer cells into induced pluripotent stem cells (iPSCs) opens up the possibility of converting malignant cells into any cell type, including those best suited to be developed as cancer vaccines. Mouse models are needed to evaluate and optimize the therapeutic efficacy of such novel cancer vaccines. However, only human cancer cell lines have been reported as being reprogrammed into iPSCs. Here, we report a proof-of-principle study which shows that mouse cancer cells can be reprogrammed into iPSCs that are capable of subsequent differentiation. Four canonical reprogramming transcription factors, Oct3/4, Sox2, Klf4, and c-Myc, were introduced by plasmid transfection into mouse Lewis lung carcinoma D122 harboring Nanog-GFP reporter. Green fluorescent cells were found clustered into embryonic stem cell (ESC)-like colonies expressing ESC markers, Oct4 and SSEA-1. Bisulfite genomic sequencing analyses of these cells revealed hypomethylation of the Nanog promoter. The expression of a host of pluripotency genes by these reprogrammed cells at levels similar to those of ESCs was confirmed by quantitative real-time PCR. Functional pluripotency of the reprogrammed cells was demonstrated by their ability to form embryoid bodies and differentiate into neuronal progenitors on retinoic acid treatment. This study indicates the feasibility of developing iPSC-based experimental cancer vaccines for immunotherapy in mouse models.
Department of Chemical Pathology and Sir Y.K. Pao Centre for Cancer, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong SAR, China.
Frances Ka-Yin
FK
Lin
Yiu-Loon
YL
Chui
D000818:Animals; D018827:Carcinoma, Lewis Lung / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002454:Cell Differentiation / Q000502:physiology; D065150:Cellular Reprogramming; D004195:Disease Models, Animal; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D000090062:Kruppel-Like Factor 4; D051379:Mice; D014162:Transfection
C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D049452:Green Fluorescent Proteins