Limbal niche cells can reduce the angiogenic potential of cultivated oral mucosal epithelial cells.

Autor: Duan CY; Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 China., Xie HT; Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 China., Zhao XY; Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 China., Xu WH; Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 China., Zhang MC; Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 China.
Jazyk: angličtina
Zdroj: Cellular & molecular biology letters [Cell Mol Biol Lett] 2019 Apr 04; Vol. 24, pp. 3. Date of Electronic Publication: 2019 Apr 04 (Print Publication: 2019).
DOI: 10.1186/s11658-018-0133-x
Abstrakt: Background: Autologous cultivated oral mucosal epithelial transplantation (COMET) is an important treatment for limbal stem cell deficiency. However, peripheral corneal neovascularization after surgery hinders its application. This study aims to employ a culture system using allogenic limbal niche cells (LNCs) instead of mouse-derived 3T3 cells as a feeder layer that could relieve postoperative neovascularization.
Methods: Rat oral mucosal epithelial cells (OMECs) were co-cultured with rat LNCs or 3T3 cells. Cultivated oral mucosal epithelial cells (COMECs) of different culture systems were identified by hematoxylin and eosin staining and immunocytochemistry. The expression levels of the angiogenesis-related factors were analyzed by RT-qPCR and western blotting/ELISA. Angiogenic potential was reconfirmed by cell viability and tube formation assays with human umbilical vein endothelial cells (HUVECs).
Results: COMECs were obtained from both culture systems successfully. Immunocytochemistry showed approximately equal percentages of positive staining cells for p63α ( p  = 0.9177), ABCG2 ( p  = 0.526), Ki67 ( p  = 0.0987), and CK3 ( p  = 0.4000) in COMECs of different groups. RT-qPCR and western blotting/ELISA showed that COMECs of the LNC group expressed a significantly lower amount of basic fibroblast growth factor (bFGF) ( p  = 0.0038 for RT-qPCR, p  = 0.0026 for western blotting) but more pigment epithelium-derived factor (PEDF) ( p  = 0.0172 for RT-qPCR, p  = 0.0253 for western blotting) and soluble fms-like tyrosine kinase-1 (sFlt-1) ( p  < 0.0001 for RT-qPCR, p  = 0.0064 for ELISA) than the COMECs of the 3T3 group. Furthermore, compared with COMECs of the 3T3 group, COMECs of the LNC group could reduce the viability ( p  = 0.0002) and tube formation ( p  = 0.0002) of HUVECs.
Conclusions: LNCs could substitute 3T3 cells for expanding OMECs in vitro, and the COMECs obtained in this system are less likely to induce postsurgical neovascularization, which provides an alternative option for an ex vivo culture system and promotes the application of COMET.
Competing Interests: No human-related experiments were performed in our study. The animal-related activities were adherent to the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research and approved by the Institutional Animal Care and Use Committee at Tongji Medical College, Huazhong University of Science and Technology (Permit Number: S641).Not applicable.The authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Databáze: MEDLINE
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