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Improvement of tube formation model of cell: Application for acute hypoxia in in vitro study of angiogenesis.
Li, Fengyang; Gou, Xue; Xu, Dan; Han, Dan; Hou, Kai; Fang, Weirong; Li, Yunman.
Afiliación
  • Li F; State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, Nanjing 210009, PR China; Department of Pharmacology, Key Laboratory of Chemical Biology (Ministry of Education), School of Pharmaceutical Sciences, Shandong University, Jinan, Shandong, China.
  • Gou X; State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, Nanjing 210009, PR China.
  • Xu D; State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, Nanjing 210009, PR China.
  • Han D; Department of Pharmacy, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing 210008, PR China.
  • Hou K; Department of Pharmacy, Zhongda Hospital, Southeast University, Nanjing 210009, PR China.
  • Fang W; State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, Nanjing 210009, PR China. Electronic address: weirongfang@163.com.
  • Li Y; State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, Nanjing 210009, PR China. Electronic address: yunmanlicpu@163.com.
Microvasc Res ; 140: 104297, 2022 03.
Article en En | MEDLINE | ID: mdl-34890690
Angiogenesis caused by acute vascular occlusion occurs in various ischemic diseases. The in vitro tube formation assay by endothelial cells is a rapid, quantitative method for drug discovery on angiogenesis. Tube formation assay on Matrigel has been widely used to identify the angiogenesis, however, there are some problems to limit its application. In this study, we found for the first time that sodium dithionite (SD) could induce endothelial cell tube formation without Matrigel under hypoxia condition. To further verify our findings, the angiogenesis related proteins and mRNA at different time points after tube formation were measured both in primary human large-vessel endothelial cell (HUVECs) and murine microvascular endothelial cell line (Bend.3). In conclusion, compared with traditional tube formation on Matrigel, the novel model exhibits the following advantages: (1) Combination oxygen glucose deprivation with sodium dithionite (OGD-SD) model is operated more easily than traditional tube formation. (2) OGD-SD can be used for not only cell imaging, but also immunofluorescence, protein extraction and gene analysis. (3) OGD-SD is more applicable to acute hypoxia model of endothelial cell in vitro. (4) OGD-SD may be more suitable to identify molecular mechanism of compound that intervenes processes of pro-tube formation, tube formation and tube disconnection.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Neovascularización Fisiológica / Células Endoteliales / Células Endoteliales de la Vena Umbilical Humana / Neovascularización Patológica Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: Microvasc Res Año: 2022 Tipo del documento: Article País de afiliación: China Pais de publicación: Estados Unidos
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Neovascularización Fisiológica / Células Endoteliales / Células Endoteliales de la Vena Umbilical Humana / Neovascularización Patológica Tipo de estudio: Prognostic_studies Límite: Animals / Humans Idioma: En Revista: Microvasc Res Año: 2022 Tipo del documento: Article País de afiliación: China Pais de publicación: Estados Unidos