RRC ID 21534
Author Sonoyama T, Sone M, Honda K, Taura D, Kojima K, Inuzuka M, Kanamoto N, Tamura N, Nakao K.
Title Differentiation of human embryonic stem cells and human induced pluripotent stem cells into steroid-producing cells.
Journal Endocrinology
Abstract Although there have been reports of the differentiation of mesenchymal stem cells and mouse embryonic stem (ES) cells into steroid-producing cells, the differentiation of human ES/induced pluripotent stem (iPS) cells into steroid-producing cells has not been reported. The purpose of our present study was to establish a method for inducing differentiation of human ES/iPS cells into steroid-producing cells. The first approach we tried was embryoid body formation and further culture on adherent plates. The resultant differentiated cells expressed mRNA encoding the steroidogenic enzymes steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase, cytochrome P450-containing enzyme (CYP)-11A1, CYP17A1, and CYP19, and secreted progesterone was detected in the cell medium. However, expression of human chorionic gonadotropin was also detected, suggesting the differentiated cells were trophoblast like. We next tried a multistep approach. As a first step, human ES/iPS cells were induced to differentiate into the mesodermal lineage. After 7 d of differentiation induced by 6-bromoindirubin-3'-oxime (a glycogen synthase kinase-3β inhibitor), the human ES/iPS cells had differentiated into fetal liver kinase-1- and platelet derived growth factor receptor-α-expressing mesodermal lineage cells. As a second step, plasmid DNA encoding steroidogenic factor-1, a master regulator of steroidogenesis, was introduced into these mesodermal cells. The forced expression of steroidogenic factor-1 and subsequent addition of 8-bromoadenosine 3',5'-cyclic monophosphate induced the mesodermal cells to differentiate into the steroidogenic cell lineage, and expression of CYP21A2 and CYP11B1, in addition to steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase, CYP11A1, and CYP17A1, was detected. Moreover, secreted cortisol was detected in the medium, but human chorionic gonadotropin was not. These findings indicate that the steroid-producing cells obtained through the described multistep method are not trophoblast like; instead, they exhibit characteristics of adrenal cortical cells.
Volume 153(9)
Pages 4336-45
Published 2012-9
DOI 10.1210/en.2012-1060
PII en.2012-1060
PMID 22778223
MeSH Blotting, Western Cell Differentiation / physiology Cell Line Embryoid Bodies / cytology Embryonic Stem Cells / cytology* Embryonic Stem Cells / metabolism* Flow Cytometry Humans Immunohistochemistry Induced Pluripotent Stem Cells / cytology* Induced Pluripotent Stem Cells / metabolism* Real-Time Polymerase Chain Reaction Steroids / metabolism*
IF 3.961
Times Cited 12
WOS Category ENDOCRINOLOGY & METABOLISM
Resource
DNA material pCMFlag_hsNR5A1 (RDB06299)