Recapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model

To ensure nutrient and oxygen supply, tumors beyond a size of 1–2 mm<sup>3</sup> need a connection to the vascular system. Thus, tumor cells modify physiological tissue homeostasis by secreting inflammatory and angiogenic cytokines. This leads to the activation of the tumor microenvironm...

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Autores principales:	Claudia Kuehlbach, Sabine Hensler, Margareta M. Mueller
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	angiogenesis tubular structures vascular sprouting high-throughput 3D tumor-stroma model tumor–stroma interaction dextran hydrogel Technology T Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/246a956929554e52aebd565abee79c04
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spelling	oai:doaj.org-article:246a956929554e52aebd565abee79c042021-11-25T16:46:41ZRecapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model10.3390/bioengineering81101862306-5354https://doaj.org/article/246a956929554e52aebd565abee79c042021-11-01T00:00:00Zhttps://www.mdpi.com/2306-5354/8/11/186https://doaj.org/toc/2306-5354To ensure nutrient and oxygen supply, tumors beyond a size of 1–2 mm<sup>3</sup> need a connection to the vascular system. Thus, tumor cells modify physiological tissue homeostasis by secreting inflammatory and angiogenic cytokines. This leads to the activation of the tumor microenvironment and the turning of the angiogenic switch, resulting in tumor vascularization and growth. To inhibit tumor growth by developing efficient anti-angiogenic therapies, an in depth understanding of the molecular mechanism initiating angiogenesis is essential. Yet so far, predominantly 2D cell cultures or animal models have been used to clarify the interactions within the tumor stroma, resulting in poor transferability of the data obtained to the in vivo situation. Consequently, there is an abundant need for complex, humanized, 3D models in vitro. We established a dextran-hydrogel-based 3D organotypic in vitro model containing microtumor spheroids, macrophages, neutrophils, fibroblasts and endothelial cells, allowing for the analysis of tumor–stroma interactions in a controlled and modifiable environment. During the cultivation period of 21 days, the microtumor spheroids in the model grew in size and endothelial cells formed elongated tubular structures resembling capillary vessels, that appeared to extend towards the tumor spheroids. The tubular structures exhibited complex bifurcations and expanded without adding external angiogenic factors such as VEGF to the culture. To allow high-throughput screening of therapeutic candidates, the 3D cell culture model was successfully miniaturized to a 96-well format, while still maintaining the same level of tumor spheroid growth and vascular sprouting. The quantification of VEGF in the conditioned medium of these cultures showed a continuous increase during the cultivation period, suggesting the contribution of endogenous VEGF to the induction of the angiogenic switch and vascular sprouting. Thus, this model is highly suitable as a testing platform for novel anticancer therapeutics targeting the tumor as well as the vascular compartment.Claudia KuehlbachSabine HenslerMargareta M. MuellerMDPI AGarticleangiogenesistubular structuresvascular sproutinghigh-throughput 3D tumor-stroma modeltumor–stroma interactiondextran hydrogelTechnologyTBiology (General)QH301-705.5ENBioengineering, Vol 8, Iss 186, p 186 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	angiogenesis tubular structures vascular sprouting high-throughput 3D tumor-stroma model tumor–stroma interaction dextran hydrogel Technology T Biology (General) QH301-705.5
spellingShingle	angiogenesis tubular structures vascular sprouting high-throughput 3D tumor-stroma model tumor–stroma interaction dextran hydrogel Technology T Biology (General) QH301-705.5 Claudia Kuehlbach Sabine Hensler Margareta M. Mueller Recapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model
description	To ensure nutrient and oxygen supply, tumors beyond a size of 1–2 mm<sup>3</sup> need a connection to the vascular system. Thus, tumor cells modify physiological tissue homeostasis by secreting inflammatory and angiogenic cytokines. This leads to the activation of the tumor microenvironment and the turning of the angiogenic switch, resulting in tumor vascularization and growth. To inhibit tumor growth by developing efficient anti-angiogenic therapies, an in depth understanding of the molecular mechanism initiating angiogenesis is essential. Yet so far, predominantly 2D cell cultures or animal models have been used to clarify the interactions within the tumor stroma, resulting in poor transferability of the data obtained to the in vivo situation. Consequently, there is an abundant need for complex, humanized, 3D models in vitro. We established a dextran-hydrogel-based 3D organotypic in vitro model containing microtumor spheroids, macrophages, neutrophils, fibroblasts and endothelial cells, allowing for the analysis of tumor–stroma interactions in a controlled and modifiable environment. During the cultivation period of 21 days, the microtumor spheroids in the model grew in size and endothelial cells formed elongated tubular structures resembling capillary vessels, that appeared to extend towards the tumor spheroids. The tubular structures exhibited complex bifurcations and expanded without adding external angiogenic factors such as VEGF to the culture. To allow high-throughput screening of therapeutic candidates, the 3D cell culture model was successfully miniaturized to a 96-well format, while still maintaining the same level of tumor spheroid growth and vascular sprouting. The quantification of VEGF in the conditioned medium of these cultures showed a continuous increase during the cultivation period, suggesting the contribution of endogenous VEGF to the induction of the angiogenic switch and vascular sprouting. Thus, this model is highly suitable as a testing platform for novel anticancer therapeutics targeting the tumor as well as the vascular compartment.
format	article
author	Claudia Kuehlbach Sabine Hensler Margareta M. Mueller
author_facet	Claudia Kuehlbach Sabine Hensler Margareta M. Mueller
author_sort	Claudia Kuehlbach
title	Recapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model
title_short	Recapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model
title_full	Recapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model
title_fullStr	Recapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model
title_full_unstemmed	Recapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model
title_sort	recapitulating the angiogenic switch in a hydrogel-based 3d in vitro tumor-stroma model
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/246a956929554e52aebd565abee79c04
work_keys_str_mv	AT claudiakuehlbach recapitulatingtheangiogenicswitchinahydrogelbased3dinvitrotumorstromamodel AT sabinehensler recapitulatingtheangiogenicswitchinahydrogelbased3dinvitrotumorstromamodel AT margaretammueller recapitulatingtheangiogenicswitchinahydrogelbased3dinvitrotumorstromamodel
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Recapitulating the Angiogenic Switch in a Hydrogel-Based 3D In Vitro Tumor-Stroma Model

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