Gene expression profiling of mouse embryonic stem cell progeny differentiated by Lumelsky's protocol.
Successful conversion of embryonic stem (ES) cells into insulin-producing cells has been reported by Lumelsky et al. (Science 2001;292:1389-1394); however, it remains controversial. In this study, we investigated the properties of ES cell progeny-induced differentiation according to Lumelsky's protocol by immunocytochemistry, oligonucleotide microarray and real-time RT-PCR. Insulin-positive cells were observed at stages 3, 4 and 5. Microarray analysis demonstrated upregulation and appearance of some genes involved in pancreatic development but not beta-cell-specific functional genes in cells at stage 5. Similarly, real-time RT-PCR revealed that expression of beta-cell-specific functional genes such as islet amyloid polypeptide, insulin I and II was not increased in cells at stage 5. These results suggest that terminal differentiation of ES cell progeny toward functional pancreatic beta-cell is insufficient. This study also demonstrates the usefulness of multiple time-course expression profiles for validating differentiation fates of ES cell progeny.
Kitano
Mami
M
Department of Biomedical Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan. aa47155@mail.ecc.u-tokyo.ac.jp
Kakinuma
Michiru
M
Takatori
Atsushi
A
Negishi
Takayuki
T
Ishii
Yoshiyuki
Y
Kyuwa
Shigeru
S
Yoshikawa
Yasuhiro
Y
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D007150:Immunohistochemistry; D007328:Insulin / Q000032:analysis / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000196:embryology / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013687:Telencephalon / Q000378:metabolism
D007328:Insulin