Long-term expansion of directly reprogrammed keratinocyte-like cells and in vitro reconstitution of human skin

Jie Zheng, Wonjin Yun, Junghyun Park, Phil Jun Kang, Gilju Lee, Gwonhwa Song, In Yong Kim, Seungkwon You

Research output: Contribution to journalArticle

Abstract

Background: Human keratinocytes and derived products are crucial for skin repair and regeneration. Despite substantial advances in engineered skin equivalents, their poor availability and immunorejection remain major challenges in skin grafting. Methods: Induced keratinocyte-like cells (iKCs) were directly reprogrammed from human urine cells by retroviral transduction of two lineage-specific transcription factors BMI1 and ΔNP63α (BN). Expression of keratinocyte stem cell or their differentiation markers were assessed by PCR, immunofluorescence and RNA-Sequencing. Regeneration capacity of iKCs were assessed by reconstitution of a human skin equivalent under air-interface condition. Results: BN-driven iKCs were similar to primary keratinocytes (pKCs) in terms of their morphology, protein expression, differentiation potential, and global gene expression. Moreover, BN-iKCs self-assembled to form stratified skin equivalents in vitro. Conclusions: This study demonstrated an approach to generate human iKCs that could be directly reprogrammed from human somatic cells and extensively expanded in serum- and feeder cell-free systems, which will facilitate their broad applicability in an efficient and patient-specific manner.

Original languageEnglish
Article number56
JournalJournal of Biomedical Science
Volume27
Issue number1
DOIs
Publication statusPublished - 2020 Apr 20

Keywords

  • Direct lineage reprogramming
  • Induced keratinocyte-like cells
  • Long-term expansion
  • Skin reconstitution
  • Urine cells

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology
  • Biochemistry, medical
  • Pharmacology (medical)

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