Transdifferentiation of rat keratinocyte progenitors to corneal epithelial cells by limbal niche via the STAT3/PI3K/AKT signaling pathway
Abstract Purpose To develop a method for enriching keratinocyte progenitor cells (KPCs) and establish a limbal niche (LN)-mediated transdifferentiation protocol of KPCs into corneal epithelial cells. Methods Limbal niche cells (LNCs) were isolated from limbal tissues through enzymatic digestion and...
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BMC
2025-01-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-024-04129-w |
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| author | Bei Wang Jiang-Lan Zhao Gong-Yue Wang Wan-Ying Cai Yu-Ting Xiao Jia-Song Wang Chao Wang Yu-Zhi Li Xi Peng Tian-Yu Yao Ming-Chang Zhang Hua-Tao Xie |
| author_facet | Bei Wang Jiang-Lan Zhao Gong-Yue Wang Wan-Ying Cai Yu-Ting Xiao Jia-Song Wang Chao Wang Yu-Zhi Li Xi Peng Tian-Yu Yao Ming-Chang Zhang Hua-Tao Xie |
| author_sort | Bei Wang |
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| description | Abstract Purpose To develop a method for enriching keratinocyte progenitor cells (KPCs) and establish a limbal niche (LN)-mediated transdifferentiation protocol of KPCs into corneal epithelial cells. Methods Limbal niche cells (LNCs) were isolated from limbal tissues through enzymatic digestion and characterized. Conditioned medium from LNCs cultures was collected. KPCs were enriched by rapid adhesion of Matrigel and subsequently cultured in either an LNCs-conditioned medium supplemented with KSFM (LN-KS) or SHEM (LN-SH) for 14 days. Corneal-specific marker expression was assessed to evaluate transdifferentiation efficiency. Key transcription factors and signaling pathways involved in the transdifferentiation process were identified through single-cell and RNA sequencing, and were validated by western blot and quantitative real-time PCR. Results Both LN-KS and LN-SH protocols successfully induced corneal epithelial cell transdifferentiation from KPCs, with LN-KS demonstrating higher efficiency in generating CK12 + and p63 + cells (p < 0.001). RNA sequencing analysis and western blot have revealed significant activation of STAT3 and PI3K/AKT signaling pathways. Inhibition of STAT3 blocked the activation of PI3K/AKT signaling pathway and impaired corneal epithelial cell transdifferentiation. Conclusions This study demonstrates the ability of LN to promote KPCs transdifferentiation into corneal epithelial cells in vitro, and this process is partially mediated by the STAT3/PI3K/AKT signaling pathway. Graphical abstract |
| format | Article |
| id | doaj-art-dbb343233ac147908526f4e351802c1f |
| institution | Kabale University |
| issn | 1757-6512 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
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| series | Stem Cell Research & Therapy |
| spelling | doaj-art-dbb343233ac147908526f4e351802c1f2025-01-12T12:10:19ZengBMCStem Cell Research & Therapy1757-65122025-01-0116111910.1186/s13287-024-04129-wTransdifferentiation of rat keratinocyte progenitors to corneal epithelial cells by limbal niche via the STAT3/PI3K/AKT signaling pathwayBei Wang0Jiang-Lan Zhao1Gong-Yue Wang2Wan-Ying Cai3Yu-Ting Xiao4Jia-Song Wang5Chao Wang6Yu-Zhi Li7Xi Peng8Tian-Yu Yao9Ming-Chang Zhang10Hua-Tao Xie11Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and TechnologyAbstract Purpose To develop a method for enriching keratinocyte progenitor cells (KPCs) and establish a limbal niche (LN)-mediated transdifferentiation protocol of KPCs into corneal epithelial cells. Methods Limbal niche cells (LNCs) were isolated from limbal tissues through enzymatic digestion and characterized. Conditioned medium from LNCs cultures was collected. KPCs were enriched by rapid adhesion of Matrigel and subsequently cultured in either an LNCs-conditioned medium supplemented with KSFM (LN-KS) or SHEM (LN-SH) for 14 days. Corneal-specific marker expression was assessed to evaluate transdifferentiation efficiency. Key transcription factors and signaling pathways involved in the transdifferentiation process were identified through single-cell and RNA sequencing, and were validated by western blot and quantitative real-time PCR. Results Both LN-KS and LN-SH protocols successfully induced corneal epithelial cell transdifferentiation from KPCs, with LN-KS demonstrating higher efficiency in generating CK12 + and p63 + cells (p < 0.001). RNA sequencing analysis and western blot have revealed significant activation of STAT3 and PI3K/AKT signaling pathways. Inhibition of STAT3 blocked the activation of PI3K/AKT signaling pathway and impaired corneal epithelial cell transdifferentiation. Conclusions This study demonstrates the ability of LN to promote KPCs transdifferentiation into corneal epithelial cells in vitro, and this process is partially mediated by the STAT3/PI3K/AKT signaling pathway. Graphical abstracthttps://doi.org/10.1186/s13287-024-04129-w Corneal epithelial cellsTransdifferentiationKeratinocyte progenitorsLimbal nicheSTAT3PI3K/AKT signaling pathway |
| spellingShingle | Bei Wang Jiang-Lan Zhao Gong-Yue Wang Wan-Ying Cai Yu-Ting Xiao Jia-Song Wang Chao Wang Yu-Zhi Li Xi Peng Tian-Yu Yao Ming-Chang Zhang Hua-Tao Xie Transdifferentiation of rat keratinocyte progenitors to corneal epithelial cells by limbal niche via the STAT3/PI3K/AKT signaling pathway Stem Cell Research & Therapy Corneal epithelial cells Transdifferentiation Keratinocyte progenitors Limbal niche STAT3 PI3K/AKT signaling pathway |
| title | Transdifferentiation of rat keratinocyte progenitors to corneal epithelial cells by limbal niche via the STAT3/PI3K/AKT signaling pathway |
| title_full | Transdifferentiation of rat keratinocyte progenitors to corneal epithelial cells by limbal niche via the STAT3/PI3K/AKT signaling pathway |
| title_fullStr | Transdifferentiation of rat keratinocyte progenitors to corneal epithelial cells by limbal niche via the STAT3/PI3K/AKT signaling pathway |
| title_full_unstemmed | Transdifferentiation of rat keratinocyte progenitors to corneal epithelial cells by limbal niche via the STAT3/PI3K/AKT signaling pathway |
| title_short | Transdifferentiation of rat keratinocyte progenitors to corneal epithelial cells by limbal niche via the STAT3/PI3K/AKT signaling pathway |
| title_sort | transdifferentiation of rat keratinocyte progenitors to corneal epithelial cells by limbal niche via the stat3 pi3k akt signaling pathway |
| topic | Corneal epithelial cells Transdifferentiation Keratinocyte progenitors Limbal niche STAT3 PI3K/AKT signaling pathway |
| url | https://doi.org/10.1186/s13287-024-04129-w |
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