International Journal of Stem Cells

  • 제16권4호
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  • Pages.394-405
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  • 2023
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  • 2005-3606(pISSN)
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  • 2005-5447(eISSN)
한국줄기세포학회 (Korean Society for Stem Cell Research)
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Differentiation and Characterization of Cystic Fibrosis Transmembrane Conductance Regulator Knockout Human Pluripotent Stem Cells into Salivary Gland Epithelial Progenitors

  • Shuang Yan (Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology and Central Laboratory, Peking University) ;
  • Yifei Zhang (Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology and Central Laboratory, Peking University) ;
  • Siqi Zhang (Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology and Central Laboratory, Peking University) ;
  • Shicheng Wei (Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology and Central Laboratory, Peking University)
  • 투고 : 2023.03.24
  • 심사 : 2023.06.24
  • 발행 : 2023.11.30
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초록

The differentiation of pluripotent stem cells has been used to study disease mechanisms and development. We previously described a method for differentiating human pluripotent stem cells (hPSCs) into salivary gland epithelial progenitors (SGEPs). Here, cystic fibrosis transmembrane conductance regulator (CFTR) knockout hPSCs were differentiated into SGEPs derived from CFTR knockout hESCs (CF-SGEPs) using the same protocol to investigate whether the hPSC-derived SGEPs can model the characteristics of CF. CF-a disease that affects salivary gland (SG) function-is caused by mutations of the CFTR gene. Firstly, we successfully generated CFTR knockout hPSCs with reduced CFTR protein expression using the CRISPR-Cas9 system. After 16 days of differentiation, the protein expression of CFTR decreased in SGEPs derived from CFTR knockout hESCs (CF-SGEPs). RNA-Seq revealed that multiple genes modulating SG development and function were down-regulated, and positive regulators of inflammation were up-regulated in CF-SGEPs, correlating with the salivary phenotype of CF patients. These results demonstrated that CFTR suppression disrupted the differentiation of hPSC-derived SGEPs, which modeled the SG development of CF patients. In summary, this study not only proved that the hPSC-derived SGEPs could serve as manipulable and readily accessible cell models for the study of SG developmental diseases but also opened up new avenues for the study of the CF mechanism.

키워드

과제정보

We thank National Center for Protein Sciences at Peking University in Beijing, China, for assistance with providing Roche qPCR equipment and Nikon A1R confocal microscopy photography.

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