Protocol for differentiation of human embryonic stem cells into surface epithelium and functional keratinocytes

Jiafeng Liu; Huaxing Huang; Hong Ouyang

doi:10.1016/j.xpro.2025.103919

Protocol for differentiation of human embryonic stem cells into surface epithelium and functional keratinocytes

STAR Protoc. 2025 Jun 27;6(3):103919. doi: 10.1016/j.xpro.2025.103919. Online ahead of print.

Authors

Jiafeng Liu¹, Huaxing Huang², Hong Ouyang³

Affiliations

¹ State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, China. Electronic address: liujf7@mail2.sysu.edu.cn.
² State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, China.
³ State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou 510060, China; Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou 510060, China. Electronic address: ouyhong3@mail.sysu.edu.cn.

PMID: 40580474
DOI: 10.1016/j.xpro.2025.103919

Abstract

The normal development and function of the epidermis and other ectodermal appendages depend on the proper formation of the surface epithelium (SE). Here, we present a protocol to differentiate human embryonic stem cells (hESCs) into SE and subsequently into epidermal keratinocyte progenitors. We then detail procedures for the passage and expansion of keratinocyte progenitors, followed by 2D validation of terminal differentiation and 3D assessment of stratification potential. This protocol represents a promising advancement for stem cell-based regenerative medicine. For complete details on the use and execution of this protocol, please refer to Liu et al.¹ and Huang et al.².

Keywords: Cell Biology; Cell culture; Stem Cells.