Regeneration of a bioengineered 3D integumentary organ system from iPS cells

Nat Protoc. 2019 May;14(5):1323-1338. doi: 10.1038/s41596-019-0124-z. Epub 2019 Apr 8.

Abstract

Organ systems play essential roles in the physiological functions required for homeostasis. A 3D integumentary organ system (3D-IOS) comprises the skin and skin appendages such as hair follicles and sebaceous glands. This protocol describes how to induce the differentiation of murine induced pluripotent stem (iPS) cells into a 3D-IOS. First, iPS cells are grown for 7 d under conditions that encourage the formation of embryoid bodies (EBs). The iPS cell-derived EBs are stimulated by Wnt10b one day before transplantation of multiple EBs in vivo (a method we describe as the clustering-dependent embryoid body (CDB) transplantation method). After a further 30 d, the transplanted EBs will have differentiated into a 3D-IOS containing mature hair follicles and sebaceous glands. These can be removed and transplanted into wounds in the skin of other mice. After transplantation of a 3D-IOS, the organ system shows full physiological function in vivo starting 14 d following transplant. Thus, this protocol enables a whole functional organ system to be generated from pluripotent stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques
  • Cell Differentiation
  • Cell Line
  • Cells, Cultured
  • Embryoid Bodies / cytology
  • Female
  • Hair Follicle / cytology*
  • Induced Pluripotent Stem Cells / cytology*
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Proto-Oncogene Proteins
  • Regeneration / physiology*
  • Sebaceous Glands / cytology*
  • Tissue Engineering / methods*
  • Wnt Proteins

Substances

  • Proto-Oncogene Proteins
  • Wnt Proteins