Open AccessGeneration 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
Author(s) -
Juliet O. Makanga,
Michisuke Kobayashi,
Hiroki Ikeda,
Antonius Christianto,
Hidenao Toyoda,
Mitsunori Yamada,
Toshisuke Kawasaki,
Toshiyuki Inazu
Publication year - 2015
Publication title -
biological and pharmaceutical bulletin
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.635
H-Index - 117
eISSN - 1347-5215
pISSN - 0918-6158
DOI - 10.1248/bpb.b14-00697
Subject(s) - induced pluripotent stem cell , sox2 , biology , epitope , clone (java method) , stem cell , microbiology and biotechnology , glycan , antibody , embryonic stem cell , immunology , genetics , gene , glycoprotein
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.