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Identification of a new way to induce differentiation of dermal fibroblasts into vascular endothelial cells
BACKGROUND: Human dermal fibroblasts (HDFs) have the potential to differentiate into vascular endothelial cells (VECs), but their differentiation rate is low and the mechanism involved is not clear. The small molecule pathway controls the phenotype of fibroblasts by activating cellular signaling pat...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9549676/ https://www.ncbi.nlm.nih.gov/pubmed/36210433 http://dx.doi.org/10.1186/s13287-022-03185-4 |
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author | Cui, XiaoLing Wang, XiaoTan Wen, Jie Li, Xiao Li, Nan Hao, XuXiao Zhao, BaoXiang Wu, Xunwei Miao, JunYing |
author_facet | Cui, XiaoLing Wang, XiaoTan Wen, Jie Li, Xiao Li, Nan Hao, XuXiao Zhao, BaoXiang Wu, Xunwei Miao, JunYing |
author_sort | Cui, XiaoLing |
collection | PubMed |
description | BACKGROUND: Human dermal fibroblasts (HDFs) have the potential to differentiate into vascular endothelial cells (VECs), but their differentiation rate is low and the mechanism involved is not clear. The small molecule pathway controls the phenotype of fibroblasts by activating cellular signaling pathways, which is a more convenient method in the differentiation strategy of HDFs into VECs. METHODS: In this study, HDFs were treated with the different doses of CPP ((E)-4-(4-(4-(7-(diethylamino)-2-oxo-2H-chromene-3-carbonyl) piperazin-1-yl) styryl)-1-methylpyridin-1-ium iodide), and the mRNA and protein levels of HDFs were detected by qPCR, Western blot, flow cytometry and immunofluorescent staining. The matrigel assays, acetylated-LDL uptake and angiogenesis assays of chick embryo chorioallantoic membrane (CAM) and hindlimb ischemia model of nude mice were performed to evaluate the functions of VECs derived from HDFs. RESULTS: Here, we report that the small chemical molecule, CPP, can effectively induce HDFs to differentiate into VECs. First, we observed the morphological changes of HDFS treated with CPP. Flow cytometry, Western blot and qRT-PCR analyses showed that CPP effectively decreased the level of the HDFs-marker Vimentin and increased levels of the VEC-markers CD31, CD133, TEK, ERG, vWF, KDR and CDH5. Detection of the percentage of CD31-positive cells by immunofluorescent staining confirmed that CPP can effectively induce HDFs to differentiate into VECs. The results of Matrigel assays, DiI-ac-LDL uptake, angiogenesis assays on CAM and hindlimb ischemia model of nude mice showed that CPP-induced HDFs have the functions of VECs in vitro and in vivo. Western blot and qRT-PCR analysis showed that CPP induces HDFs to differentiate into VECs by promoting the expression of pro-angiogenic factors (VEGF, FGF-2 and PDGF-BB). CONCLUSIONS: Our data suggest that the small chemical molecule CPP efficiently induces the differentiation of HDFs into VECs. Simultaneously, this new inducer provides a potential to develop new approaches to restore vascular function for the treatment of ischemic vascular diseases. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-022-03185-4. |
format | Online Article Text |
id | pubmed-9549676 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-95496762022-10-11 Identification of a new way to induce differentiation of dermal fibroblasts into vascular endothelial cells Cui, XiaoLing Wang, XiaoTan Wen, Jie Li, Xiao Li, Nan Hao, XuXiao Zhao, BaoXiang Wu, Xunwei Miao, JunYing Stem Cell Res Ther Research BACKGROUND: Human dermal fibroblasts (HDFs) have the potential to differentiate into vascular endothelial cells (VECs), but their differentiation rate is low and the mechanism involved is not clear. The small molecule pathway controls the phenotype of fibroblasts by activating cellular signaling pathways, which is a more convenient method in the differentiation strategy of HDFs into VECs. METHODS: In this study, HDFs were treated with the different doses of CPP ((E)-4-(4-(4-(7-(diethylamino)-2-oxo-2H-chromene-3-carbonyl) piperazin-1-yl) styryl)-1-methylpyridin-1-ium iodide), and the mRNA and protein levels of HDFs were detected by qPCR, Western blot, flow cytometry and immunofluorescent staining. The matrigel assays, acetylated-LDL uptake and angiogenesis assays of chick embryo chorioallantoic membrane (CAM) and hindlimb ischemia model of nude mice were performed to evaluate the functions of VECs derived from HDFs. RESULTS: Here, we report that the small chemical molecule, CPP, can effectively induce HDFs to differentiate into VECs. First, we observed the morphological changes of HDFS treated with CPP. Flow cytometry, Western blot and qRT-PCR analyses showed that CPP effectively decreased the level of the HDFs-marker Vimentin and increased levels of the VEC-markers CD31, CD133, TEK, ERG, vWF, KDR and CDH5. Detection of the percentage of CD31-positive cells by immunofluorescent staining confirmed that CPP can effectively induce HDFs to differentiate into VECs. The results of Matrigel assays, DiI-ac-LDL uptake, angiogenesis assays on CAM and hindlimb ischemia model of nude mice showed that CPP-induced HDFs have the functions of VECs in vitro and in vivo. Western blot and qRT-PCR analysis showed that CPP induces HDFs to differentiate into VECs by promoting the expression of pro-angiogenic factors (VEGF, FGF-2 and PDGF-BB). CONCLUSIONS: Our data suggest that the small chemical molecule CPP efficiently induces the differentiation of HDFs into VECs. Simultaneously, this new inducer provides a potential to develop new approaches to restore vascular function for the treatment of ischemic vascular diseases. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-022-03185-4. BioMed Central 2022-10-09 /pmc/articles/PMC9549676/ /pubmed/36210433 http://dx.doi.org/10.1186/s13287-022-03185-4 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Cui, XiaoLing Wang, XiaoTan Wen, Jie Li, Xiao Li, Nan Hao, XuXiao Zhao, BaoXiang Wu, Xunwei Miao, JunYing Identification of a new way to induce differentiation of dermal fibroblasts into vascular endothelial cells |
title | Identification of a new way to induce differentiation of dermal fibroblasts into vascular endothelial cells |
title_full | Identification of a new way to induce differentiation of dermal fibroblasts into vascular endothelial cells |
title_fullStr | Identification of a new way to induce differentiation of dermal fibroblasts into vascular endothelial cells |
title_full_unstemmed | Identification of a new way to induce differentiation of dermal fibroblasts into vascular endothelial cells |
title_short | Identification of a new way to induce differentiation of dermal fibroblasts into vascular endothelial cells |
title_sort | identification of a new way to induce differentiation of dermal fibroblasts into vascular endothelial cells |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9549676/ https://www.ncbi.nlm.nih.gov/pubmed/36210433 http://dx.doi.org/10.1186/s13287-022-03185-4 |
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