人羊膜上皮细胞向角膜上皮分化过程中 调控机制的研究进展
Research Advances in the Regulatory Mechanisms of Human Amniotic Epithelial Cell Differentiation into Corneal Epithelial Cells
摘要: 角膜缘干细胞缺乏症(limbal stem cell deficiency, LSCD)可破坏角膜上皮稳态，导致角膜结膜化、新生血管形成、慢性炎症及视功能损害；现有角膜缘移植虽可重建眼表，但仍受供体来源、健眼损伤风险、同种异体免疫排斥及长期免疫抑制等因素限制。人羊膜上皮细胞(human amniotic epithelial cells, hAECs)来源相对充足，具有低免疫原性、免疫调节、旁分泌活性及较低致瘤风险，并具备上皮谱系特征和多向分化潜能，是角膜上皮再生研究中具有潜力的替代种子细胞。已有体外诱导和动物实验提示hAECs可获得角膜上皮样表型并参与眼表修复，但其分化效率、表型稳定性及功能成熟仍依赖复杂的分子调控和微环境支持。本文围绕hAECs向角膜上皮方向分化的研究进展，重点综述Wnt/β-catenin、TGF-β/BMP、p38MAPK、STAT3/PI3K/AKT、EGFR/MAPK等信号通路，PAX6及p63在诱导分化与功能成熟中的作用，并进一步系统总结TEER、屏障通透性实验、细胞迁移/创面愈合实验及体内移植模型等功能性评估方法，分析功能结果与CK3/CK12、PAX6、p63、E-cadherin、ZO-1等分子标志物之间的对应关系，以期为基于hAECs的组织工程角膜及眼表再生治疗研究提供参考。
Abstract: Limbal stem cell deficiency (LSCD) disrupts corneal epithelial homeostasis and may lead to conjunctivalization of the cornea, neovascularization, chronic inflammation, and visual dysfunction. Although current limbal transplantation strategies can restore the ocular surface to some extent, their clinical application remains limited by donor tissue availability, the risk of injury to the contralateral healthy eye, immune rejection after allogeneic transplantation, and the need for long-term immunosuppression. Human amniotic epithelial cells (hAECs) are considered a promising alternative cell source for corneal epithelial regeneration because of their abundant availability, low immunogenicity, immunomodulatory capacity, paracrine activity, relatively low tumorigenic risk, epithelial lineage characteristics, and multilineage differentiation potential. Existing in vitro induction studies and animal experiments have shown that hAECs can acquire corneal epithelial-like phenotypes and contribute to ocular surface repair. However, their differentiation efficiency, phenotypic stability, and functional maturation are governed by complex molecular regulatory networks and microenvironmental cues. This review summarizes recent advances in the differentiation of hAECs toward a corneal epithelial lineage, with particular emphasis on the roles of Wnt/β-catenin, TGF-β/BMP, p38MAPK, STAT3/PI3K/AKT, and EGFR/MAPK signaling pathways, as well as PAX6 and p63 in promoting lineage specification and functional maturation, and systematically discusses functional assessment methods for hAEC-derived corneal epithelium, including transepithelial electrical resistance (TEER), barrier permeability assays, cell migration/wound-healing assays, and in vivo transplantation models, and relates functional outcomes to molecular markers such as CK3/CK12, PAX6, p63, E-cadherin, and ZO-1. This review aims to provide a theoretical reference for the development of hAEC-based tissue-engineered corneas and regenerative therapies for ocular surface diseases.
文章引用：李令洁, 梁媛婷, 刘小勇. 人羊膜上皮细胞向角膜上皮分化过程中 调控机制的研究进展[J]. 临床医学进展, 2026, 16(6): 1336-1343. https://doi.org/10.12677/acm.2026.1662344

参考文献

[1] Sun, T., Tseng, S.C. and Lavker, R.M. (2010) Location of Corneal Epithelial Stem Cells. Nature, 463, E10-E11. [Google Scholar] [CrossRef] [PubMed]
[2] Lent-Schochet, D., Akbar, M., Hou, J.H. and Farooq, A.V. (2025) Diagnostic Approach to Limbal Stem Cell Deficiency. Frontiers in Ophthalmology, 4, Article 1524595. [Google Scholar] [CrossRef] [PubMed]
[3] Jurkunas, U.V., Kaufman, A.R., Yin, J., Ayala, A., Maguire, M., Samarakoon, L., et al. (2025) Cultivated Autologous Limbal Epithelial Cell (CALEC) Transplantation for Limbal Stem Cell Deficiency: A Phase I/II Clinical Trial of the First Xenobiotic-Free, Serum-Free, Antibiotic-Free Manufacturing Protocol Developed in the US. Nature Communications, 16, Article No. 1607. [Google Scholar] [CrossRef] [PubMed]
[4] Tsai, R.J., Li, L.M. and Chen, J.K. (2000) Reconstruction of Damaged Corneas by Transplantation of Autologous Limbal Epithelial Cells. New England Journal of Medicine, 343, 86-93. [Google Scholar] [CrossRef] [PubMed]
[5] 王璐璐, 赖东梅. 人羊膜上皮细胞在妇产科相关疾病中的应用及研究进展[J]. 复旦学报(医学版), 2021, 48(6): 819-826.
[6] Zhang, Q. and Lai, D. (2020) Application of Human Amniotic Epithelial Cells in Regenerative Medicine: A Systematic Review. Stem Cell Research & Therapy, 11, Article No. 439. [Google Scholar] [CrossRef] [PubMed]
[7] Bejaoui, M., Ferdousi, F., Zheng, Y., Oda, T. and Isoda, H. (2021) Regulating Cell Fate of Human Amnion Epithelial Cells Using Natural Compounds: An Example of Enhanced Neural and Pigment Differentiation by 3,4,5-Tri-O-Caffeoylquinic Acid. Cell Communication and Signaling, 19, Article No. 26. [Google Scholar] [CrossRef] [PubMed]
[8] Michalik, M., Wieczorek, P. and Czekaj, P. (2022) In Vitro Differentiation of Human Amniotic Epithelial Cells into Hepatocyte-Like Cells. Cells, 11, Article 2138. [Google Scholar] [CrossRef] [PubMed]
[9] Zhou, Q., Liu, X., Ruan, Y., Wang, L., Jiang, M., Wu, J., et al. (2015) Construction of Corneal Epithelium with Human Amniotic Epithelial Cells and Repair of Limbal Deficiency in Rabbit Models. Human Cell, 28, 22-36. [Google Scholar] [CrossRef] [PubMed]
[10] Hu, S., Chen, J., Jin, J., Liu, Y., Xu, G. and Ou, Q. (2024) Construction of Living-Cell Tissue Engineered Amniotic Membrane for Ocular Surface Disease. BMC Ophthalmology, 24, Article No. 409. [Google Scholar] [CrossRef] [PubMed]
[11] Nguyen, H.T., Theerakittayakorn, K., Somredngan, S., Ngernsoungnern, A., Ngernsoungnern, P., Sritangos, P., et al. (2022) Signaling Pathways Impact on Induction of Corneal Epithelial-Like Cells Derived from Human Wharton’s Jelly Mesenchymal Stem Cells. International Journal of Molecular Sciences, 23, Article No. 3078. [Google Scholar] [CrossRef] [PubMed]
[12] Jurkunas, U., Johns, L. and Armant, M. (2022) Cultivated Autologous Limbal Epithelial Cell Transplantation: New Frontier in the Treatment of Limbal Stem Cell Deficiency. American Journal of Ophthalmology, 239, 244-268. [Google Scholar] [CrossRef] [PubMed]
[13] Nishida, K., Yamato, M., Hayashida, Y., Watanabe, K., Yamamoto, K., Adachi, E., et al. (2004) Corneal Reconstruction with Tissue-Engineered Cell Sheets Composed of Autologous Oral Mucosal Epithelium. New England Journal of Medicine, 351, 1187-1196. [Google Scholar] [CrossRef] [PubMed]
[14] Miki, T. (2018) Stem Cell Characteristics and the Therapeutic Potential of Amniotic Epithelial Cells. American Journal of Reproductive Immunology, 80, e13003. [Google Scholar] [CrossRef] [PubMed]
[15] García-López, G., García-Castro, I.L., Avila-González, D., et al. (2015) Las células del epitelio amniótico humano (CEAh) como posible fuente de células troncales (CT). Gaceta Médica de México, 151, 66-74.
[16] Yu, C., Yuan, H., Xu, Y., Luo, Y., Wu, Z., Zhong, J., et al. (2024) Hyaluronan Delays Human Amniotic Epithelial Stem Cell Senescence by Regulating CD44 Isoform Switch to Activate AKT/mTOR Signals. Biomedicine & Pharmacotherapy, 170, Article ID: 116100. [Google Scholar] [CrossRef] [PubMed]
[17] Yao, M., Chen, J., Yang, X.X., et al. (2013) Differentiation of Human Amniotic Epithelial Cells into Corneal Epithelial-like Cells in Vitro. International Journal of Ophthalmology, 6, 564-572.
[18] Smits, J.G.A., Cunha, D.L., Amini, M., Bertolin, M., Laberthonnière, C., Qu, J., et al. (2023) Identification of the Regulatory Circuit Governing Corneal Epithelial Fate Determination and Disease. PLOS Biology, 21, e3002336. [Google Scholar] [CrossRef] [PubMed]
[19] Stachon, T., Fries, F.N., Li, Z., Daas, L., Nagy, Z.Z., Seitz, B., et al. (2025) Decreased PAX6 and DSG1 Protein Expression in Corneal Epithelium of Patients with Epithelial Basal Membrane Dystrophy, Salzmann Nodular Degeneration, and Pterygium. Journal of Clinical Medicine, 14, Article No. 1456. [Google Scholar] [CrossRef] [PubMed]
[20] Mao, Y., Ou, S., Zhu, C., Lin, S., Liu, X., Liang, M., et al. (2022) Downregulation of p38 MAPK Signaling Pathway Ameliorates Tissue-Engineered Corneal Epithelium. Tissue Engineering Part A, 28, 977-989. [Google Scholar] [CrossRef] [PubMed]
[21] Rama, P., Matuska, S., Paganoni, G., Spinelli, A., De Luca, M. and Pellegrini, G. (2010) Limbal Stem-Cell Therapy and Long-Term Corneal Regeneration. New England Journal of Medicine, 363, 147-155. [Google Scholar] [CrossRef] [PubMed]
[22] Wang, B., Zhao, J., Wang, G., Cai, W., Xiao, Y., Wang, J., et al. (2025) Transdifferentiation of Rat Keratinocyte Progenitors to Corneal Epithelial Cells by Limbal Niche via the STAT3/PI3K/AKT Signaling Pathway. Stem Cell Research & Therapy, 16, Article No. 8. [Google Scholar] [CrossRef] [PubMed]
[23] Wang, Y., Ge, H., Chen, P. and Wang, Y. (2024) Wnt/β-Catenin Signaling in Corneal Epithelium Development, Homeostasis, and Pathobiology. Experimental Eye Research, 246, Article ID: 110022. [Google Scholar] [CrossRef] [PubMed]
[24] Shen, Y., Wang, J., Dai, Y., Wan, X., Zhang, J. and Le, Q. (2025) RSPO3 Promotes Proliferation and Self-Renewal of Limbal Epithelial Stem Cells through a WNT/β-Catenin-Independent Signaling Pathway. Investigative Ophthalmology & Visual Science, 66, Article No. 8. [Google Scholar] [CrossRef] [PubMed]
[25] Guo, Y., Wu, W., Chen, H., Wang, X., Zhang, Y., Li, S., et al. (2025) Network Analysis Reveals Potential Mechanisms That Determine the Cellular Identity of Keratinocytes and Corneal Epithelial Cells through the Hox/Gtl2-Dio3 miRNA Axis. Frontiers in Cell and Developmental Biology, 13, Article 1475334. [Google Scholar] [CrossRef] [PubMed]
[26] Dan, J., Tan, T., Wu, M., Gong, J., Yang, Q., Wang, L., et al. (2023) Lithium Chloride Promotes Diabetic Corneal Epithelial Wound Healing by Activating the Wnt/β-Catenin Signaling Pathway. Experimental and Therapeutic Medicine, 26, Article No. 373. [Google Scholar] [CrossRef] [PubMed]
[27] Wang, J., Zhu, R., Dong, Y., Feng, J., Gao, J., Wang, S., et al. (2025) YTHDF3 Suppresses Declines in Corneal Epithelial Wound Healing through Upregulating the Wnt/β-Catenin Signaling Pathway and THBS2 Expression in Diabetic Corneas. International Journal of Biological Macromolecules, 285, Article ID: 138267. [Google Scholar] [CrossRef] [PubMed]
[28] Lin, S., Cai, M., Zhang, L., Mao, Y., Wu, H., Liu, X., et al. (2023) Limbal Stem Cell Dysfunction Induced by Severe Dry Eye via Activation of the p38 MAPK Signaling Pathway. The American Journal of Pathology, 193, 1863-1878. [Google Scholar] [CrossRef] [PubMed]
[29] 胡敏, 申迪, 张美华, 等. 中性粒细胞胞外诱捕网对人羊膜上皮细胞增殖和凋亡的影响及其可能机制[J]. 中华围产医学杂志, 2022, 25(8): 609-614.
[30] 宋菲, 董叫云, 等. 人羊膜上皮细胞外泌体通过PI3K/AKT/mTOR介导血管生成和促进成纤维细胞功能加速糖尿病创面愈合[J]. 中华烧伤与创面修复杂志, 2021, 37(5): 452.
[31] Maugeri, G., D’Amico, A.G., Magrì, B., Giunta, S., Saccone, S., Federico, C., et al. (2023) Protective Effect of Pituitary Adenylate Cyclase Activating Polypeptide in Diabetic Keratopathy. Peptides, 170, Article ID: 171107. [Google Scholar] [CrossRef] [PubMed]
[32] Kao, W.W.Y., Zhang, J., Venkatakrishnan, J., Chang, S., Yuan, Y., Yamanaka, O., et al. (2024) Lumican/Lumikine Promotes Healing of Corneal Epithelium Debridement by Upregulation of EGFR Ligand Expression via Noncanonical Smad-Independent TGFβ/TBRs Signaling. Cells, 13, Article No. 1599. [Google Scholar] [CrossRef] [PubMed]
[33] Sang, T., Wang, Y., Wang, Z., Sun, D., Dou, S., Yu, Y., et al. (2024) NEAT1 Deficiency Promotes Corneal Epithelial Wound Healing by Activating cAMP Signaling Pathway. Investigative Opthalmology & Visual Science, 65, Article No. 10. [Google Scholar] [CrossRef] [PubMed]
[34] Zhu, L., Wang, L., Liu, D., Chen, C., Mo, K., Lan, X., et al. (2023) Single-Cell Transcriptomics Implicates the FEZ1-DKK1 Axis in the Regulation of Corneal Epithelial Cell Proliferation and Senescence. Cell Proliferation, 56, e13433. [Google Scholar] [CrossRef] [PubMed]
[35] Srinivasan, B., Kolli, A.R., Esch, M.B., Abaci, H.E., Shuler, M.L. and Hickman, J.J. (2015) TEER Measurement Techniques for in Vitro Barrier Model Systems. SLAS Technology, 20, 107-126. [Google Scholar] [CrossRef] [PubMed]
[36] McCullough, S. and Farber, S. (2020) Method: FITC-Dextran Trans-Epithelial Permeability Assay. Nature Publishing.
[37] Deng, S.X., Borderie, V., Chan, C.C., Dana, R., Figueiredo, F.C., Gomes, J.A.P., et al. (2018) Global Consensus on Definition, Classification, Diagnosis, and Staging of Limbal Stem Cell Deficiency. Cornea, 38, 364-375. [Google Scholar] [CrossRef] [PubMed]
[38] Ban, Y., Dota, A., Cooper, L.J., Fullwood, N.J., Nakamura, T., Tsuzuki, M., et al. (2003) Tight Junction-Related Protein Expression and Distribution in Human Corneal Epithelium. Experimental Eye Research, 76, 663-669. [Google Scholar] [CrossRef] [PubMed]
[39] Ko, J., Yanai, R. and Nishida, T. (2009) Up-Regulation of ZO-1 Expression and Barrier Function in Cultured Human Corneal Epithelial Cells by Substance P. FEBS Letters, 583, 2148-2153. [Google Scholar] [CrossRef] [PubMed]
[40] Xu, Y., Liu, J., Song, W., Wang, Q., Sun, X., Zhao, Q., et al. (2023) Biomimetic Convex Implant for Corneal Regeneration through 3D Printing. Advanced Science, 10, e2205878. [Google Scholar] [CrossRef] [PubMed]

为你推荐