title: Differentiation of human pluripotent stem cells into nephron progenitor cells in a serum and feeder free system.
abstract: OBJECTIVE
  Kidney disease is emerging as a critical medical problem worldwide. Because of limited treatment options for the damaged kidney, stem cell treatment is becoming an alternative therapeutic approach. Of many possible human stem cell sources, pluripotent stem cells are most attractive due to their self-renewal and pluripotent capacity. However, little is known about the derivation of renal lineage cells from human pluripotent stem cells (hPSCs). In this study, we developed a novel protocol for differentiation of nephron progenitor cells (NPCs) from hPSCs in a serum- and feeder-free system.
  
  
  METHODS
  We designed step-wise protocols for differentiation of human pluripotent stem cells toward primitive streak, intermediate mesoderm and NPCs by recapitulating normal nephrogenesis. Expression of key marker genes was examined by RT-PCR, real time RT-PCR and immunocytochemistry. Each experiment was independently performed three times to confirm its reproducibility.
  
  
  RESULTS
  After modification of culture period and concentration of exogenous factors, hPSCs can differentiate into NPCs that markedly express specific marker genes such as SIX2, GDNF, HOXD11, WT1 and CITED1 in addition to OSR1, PAX2, SALL1 and EYA1. Moreover, NPCs possess the potential of bidirectional differentiation into both renal tubular epithelial cells and glomerular podocytes in defined culture conditions. In particular, approximately 70% of SYN-positive cells were obtained from hPSC-derived NPCs after podocytes induction. NPCs can also form in vitro tubule-like structures in three dimensional culture systems.
  
  
  CONCLUSIONS
  Our novel protocol for hPSCs differentiation into NPCs can be useful for producing alternative sources of cell replacement therapy and disease modeling for human kidney diseases.
authors.affiliation: Graduate Schools of Medical Science and Engineering, KAIST, Daejeon, Republic of Korea.
authors.forename: Minyong
authors.initials: M
authors.lastname: Kang
authors.affiliation: Graduate Schools of Medical Science and Engineering, KAIST, Daejeon, Republic of Korea; Department of Biological Sciences, KAIST, Daejeon, Republic of Korea.
authors.forename: Yong-Mahn
authors.initials: YM
authors.lastname: Han
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology; D006801:Humans; D057026:Induced Pluripotent Stem Cells; D007684:Kidney Tubules / Q000166:cytology; D008648:Mesoderm / Q000166:cytology; D009399:Nephrons / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology*; D050199:Podocytes / Q000166:cytology; D054240:Primitive Streak / Q000166:cytology; D013234:Stem Cells / Q000166:cytology*
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