Generation of rat induced pluripotent stem cells using a plasmid vector and possible application of a keratan sulfate glycan recognizing antibody in discriminating teratoma formation phenotypes.
Induced pluripotent stem cells (iPSCs) offer an invaluable tool for biological research and regenerative medicine. We report establishment of rat iPSCs (riPSCs) using a plasmid vector encoding four transcription factors, Oct3/4, Sox2, c-Myc and Klf4. Although all riPSC clones were generated and cultured under the same conditions, expressed hallmark pluripotency markers and differentiated successfully in vitro, the expression of a keratan sulfate glycan epitope with unique properties defined by R-10G antibody varied in the riPSC clones. In contrast, tumor rejection antigen (TRA)-1-81 epitope expression was comparable. A clone highly reactive to R-10G antibody formed teratomas in vivo consisting of cells from all three germ layers. However, clones expressing a lower level of the epitope defined by R-10G resulted in tumors with rapid growth consisting of undifferentiated cells. Additionally, riPSCs could be successfully differentiated into a neuronal lineage including glutamate neurons that responded to agonist stimulation. These observations demonstrate a glycophenotypic difference that may potentially serve as a useful probe for riPSC evaluation and to study the role of glycans in pluripotency and carcinogenesis in these cells.
Laboratory of Functional Genomics, College of Pharmaceutical Sciences, Ritsumeikan University.
Juliet O
JO
Makanga
Misa
M
Kobayashi
Hiroki
H
Ikeda
Antonius
A
Christianto
Hidenao
H
Toyoda
Mitsunori
M
Yamada
Toshisuke
T
Kawasaki
Tetsuya
T
Inazu
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology; D002454:Cell Differentiation; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D007632:Keratan Sulfate / Q000276:immunology*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism; D008297:Male; D008807:Mice, Inbred BALB C; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D010641:Phenotype; D010957:Plasmids; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D017208:Rats, Wistar; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D013724:Teratoma
D000906:Antibodies; D000954:Antigens, Surface; C000715468:Klf4 protein, mouse; C000715469:Klf4 protein, rat; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D050814:Octamer Transcription Factor-3; D016271:Proto-Oncogene Proteins c-myc; D055748:SOXB1 Transcription Factors; C527784:Sox2 protein, rat; D007632:Keratan Sulfate