OBJECTIVES: Despite an increasing research demand for human microvascular endothelial cells, isolation of primary endothelial cells from human tissue remains difficult. The omentum, a highly vascular visceral adipose tissue, could provide an excellent source of these cells. METHODS: A reliable method to isolate HOMECs has been developed. It consists of initial enzymatic digestion (to deplete cell contaminants), followed by further digestion, selective filtration, and immunoselection using Dynabeads coated with CD31 antibody. Cultures were characterized for expression of endothelial cell markers and their ability to undergo VEGF-dependent in vitro tube structure formation. RESULTS: Omental-derived cultures of microvascular endothelial cells were achieved with <5% contamination of other cell types. The endothelial origin of cells was confirmed by the constitutive expression of a range of vascular endothelial markers (CD31, CD105, vWF) and internalization of DiI-AcLDL. Furthermore, cultures were negative for lymphatic endothelial markers, underwent in vitro angiogenesis, and exhibited typical endothelial morphology. CONCLUSIONS: This isolation method produces homogeneous HOMEC cultures that can be maintained in vitro for at least six passages without loss of cellular features characterizing endothelial cells.
OBJECTIVES: Despite an increasing research demand for human microvascular endothelial cells, isolation of primary endothelial cells from human tissue remains difficult. The omentum, a highly vascular visceral adipose tissue, could provide an excellent source of these cells. METHODS: A reliable method to isolate HOMECs has been developed. It consists of initial enzymatic digestion (to deplete cell contaminants), followed by further digestion, selective filtration, and immunoselection using Dynabeads coated with CD31 antibody. Cultures were characterized for expression of endothelial cell markers and their ability to undergo VEGF-dependent in vitro tube structure formation. RESULTS: Omental-derived cultures of microvascular endothelial cells were achieved with <5% contamination of other cell types. The endothelial origin of cells was confirmed by the constitutive expression of a range of vascular endothelial markers (CD31, CD105, vWF) and internalization of DiI-AcLDL. Furthermore, cultures were negative for lymphatic endothelial markers, underwent in vitro angiogenesis, and exhibited typical endothelial morphology. CONCLUSIONS: This isolation method produces homogeneous HOMEC cultures that can be maintained in vitro for at least six passages without loss of cellular features characterizing endothelial cells.
Authors: Irina Zakharova; Shoraan Saaya; Alexander Shevchenko; Alena Stupnikova; Maria Zhiven'; Pavel Laktionov; Alena Stepanova; Alexander Romashchenko; Lyudmila Yanshole; Alexander Chernonosov; Alexander Volkov; Elena Kizilova; Evgenii Zavjalov; Alexander Chernyavsky; Alexander Romanov; Andrey Karpenko; Suren Zakian Journal: Front Bioeng Biotechnol Date: 2022-03-11
Authors: Lijun Ma; Gregory S Shelness; James A Snipes; Mariana Murea; Peter A Antinozzi; Dongmei Cheng; Moin A Saleem; Simon C Satchell; Bernhard Banas; Peter W Mathieson; Matthias Kretzler; Ashok K Hemal; Lawrence L Rudel; Snezana Petrovic; Allison Weckerle; Martin R Pollak; Michael D Ross; John S Parks; Barry I Freedman Journal: J Am Soc Nephrol Date: 2014-07-10 Impact factor: 10.121
Authors: I S Zakharova; M K Zhiven'; Sh B Saaya; A I Shevchenko; A M Smirnova; A Strunov; A A Karpenko; E A Pokushalov; L N Ivanova; P I Makarevich; Y V Parfyonova; E Aboian; S M Zakian Journal: J Transl Med Date: 2017-03-03 Impact factor: 5.531
Authors: Md Zahidul I Pranjol; Dmitry A Zinovkin; Annelie R T Maskell; Laura J Stephens; Sergey L Achinovich; Dmitry M Los'; Eldar A Nadyrov; Michael Hannemann; Nicholas J Gutowski; Jacqueline L Whatmore Journal: J Transl Med Date: 2019-07-03 Impact factor: 5.531
Authors: Wei Zhang; Marc Freichel; Frank van der Hoeven; Peter Paul Nawroth; Hugo Katus; Florian Kälble; Edgar Zitron; Vedat Schwenger Journal: PLoS One Date: 2016-01-13 Impact factor: 3.240