Static and dynamic culture of human endothelial cells encapsulated inside alginate-gelatin microspheres.

Autor: Ghaleh AS; Department of Genetic, Faculty of Basic Sciences, Bonab Branch, Islamic Azad University, Bonab, Iran., Saghati S; Department of Tissue Engineering, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran., Rahbarghazi R; Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran. Electronic address: rahbarghazir@tbzmed.ac.ir., Hassani A; Chemical Engineering Faculty, Sahand University of Technology, Tabriz 51335-1996, Iran., Kaleybar LS; Chemical Engineering Faculty, Sahand University of Technology, Tabriz 51335-1996, Iran., Geranmayeh MH; Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran., Hassanpour M; Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran., Rezaie J; Solid Tumor Research Center, Cellular and Molecular Medicine Institute, Urmia University of Medical Sciences, Urmia, Iran., Soltanzadeh H; Maragheh University of Medical Sciences, Maragheh, Iran.
Jazyk: angličtina
Zdroj: Microvascular research [Microvasc Res] 2021 Sep; Vol. 137, pp. 104174. Date of Electronic Publication: 2021 May 08.
DOI: 10.1016/j.mvr.2021.104174
Abstrakt: This study aimed to explore the angiogenesis potential of human endothelial cells encapsulated inside alginate-gelatin microspheres under static and dynamic culture systems after 7 days. Human umbilical vein endothelial cells were encapsulated inside alginate (1%) and gelatin (1.2%) using an electrostatic encapsulation method. Cells were incubated for 7 days in vitro. The cell survival rate was measured using the MTT assay. The expression of VEGFR-2 and von Willebrand factor genes was studied by real-time PCR assay. Using western blot analysis, we monitored the protein contents of VEGFR-2, vWF, and Caspase 3. The levels of SOD and GPx enzymes were calculated using biochemical kits. Angiogenesis potential was assessed using in vitro Matrigel assay. Data showed an increased survival rate in encapsulated cells cultured under the static condition compared to the conventional 2D condition (p < 0.05). The culture of encapsulated cells under a dynamic bioreactor system did not alter cell viability. Compared to the dynamic culture system, the incubation of encapsulated cells in the static culture system swelled the microspheres (p < 0.05). Both dynamic and static culture models increased the expression of VEGFR-2 and von Willebrand factor in encapsulated cells compared to 2D culture (p < 0.05), showing enhanced functional maturation. Data showed a significant increase of vWF and reduction of apoptosis marker Caspase in the dynamic culture system (p < 0.05). The levels of SOD and GPx were significantly increased in dynamic and static culture models as compared to the control 2D group (p < 0.05). In vitro tubulogenesis assay showed significant induction of angiogenesis in dynamic encapsulated HUVECs indicated with a large number of vascular tubes and arborized ECs compared to the control and static encapsulated HUVECs (p < 0.05). The current study suggests a bioreactor dynamic system is a reliable approach, similar to a static condition, for the expansion of encapsulated human ECs in a 3D milieu.
(Copyright © 2021 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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