Potential of human induced pluripotent stem cells differentiating into corneal epithelial cells in simulated limbal stem cell microenvironment
10.3980/j.issn.1672-5123.2024.1.06
- VernacularTitle:模拟角膜缘干细胞微环境诱导人多潜能干细胞分化为角膜上皮细胞的研究
- Author:
Xiaofei WU
1
;
Yi ZHANG
1
;
Hongmin KE
1
;
Li CAI
1
Author Information
1. Department of Ophthalmology, Shenzhen University General Hospital, Shenzhen 518055, Guangdong Province, China
- Publication Type:Journal Article
- Keywords:
human induced pluripotent stem cells;
corneal epithelial cells;
induced differentiation
- From:
International Eye Science
2024;24(1):30-35
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To investigate the potential of human induced pluripotent stem cells(hiPSCs)differentiating into corneal epithelial cells in the simulated limbal stem cells(LSCs)microenvironment.METHODS: The hiPSC cell lines were established in vitro, and hiPSCs were co-cultured with corneal stromal cells in transwell system, which simulated the LSC microenvironment. Bone morphogenetic protein 4(BMP4)and a specific transforming growth factor β inhibitor(SB431542)were added to improve the differentiation efficacy. The expression of corneal epithelial cell-specific markers CK3 and CK12, corneal epithelial cell precursor CK15, and the limbal stem cell markers ABCG5 were determined by immunofluorescence staining and flow cytometry.RESULTS: The hiPSCs were actively proliferated in vitro, and immunofluorescence staining showed positive stem cell-specific markers OCT4, SOX2, TRA-1-60 and NANOG. Furthermore, hiPSCs co-cultured with corneal stromal cells exhibited LSCs markers ABCG5 and corneal epithelial cell precursor markers CK15 were positive; however, corneal epithelial cell markers CK3 and CK12 were negative. With the addition of BMP4 and SB431542, hiPSCs showed positive expression of CK3, and the CK3 expression increased over the time.CONCLUSION: With the addition of SB431542 and BMP4, hiPSCs cultured in simulated LSCs microenvironment could differentiate into corneal epithelial cells.
