Long-lived human lymphatic endothelial cells to study lymphatic biology and lymphatic vessel/tumor coculture in a 3D microfluidic model
Frenkel, Nicola; Poghosyan, Susanna; Alarcón, Carmen Rubio; García, Silvia Bonilla; Queiroz, Karla; Van Den Bent, Lotte; Laoukili, Jamila; Rinkes, Inne Borel; Vulto, Paul; Kranenburg, Onno; Hagendoorn, Jeroen
(2021) ACS Biomaterials Science and Engineering, volume 7, issue 7, pp. 3030 - 3042
(Article)
Abstract
The lymphatic system is essential in maintaining tissue fluid homeostasis as well as antigen and immune cell transport to lymph nodes. Moreover, lymphatic vasculature plays an important role in various pathological processes, such as cancer. Fundamental to this research field are representative in vitro models. Here we present a microfluidic
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lymphatic vessel model to study lymphangiogenesis and its interaction with colon cancer organoids using a newly developed lymphatic endothelial cell (LEC) line. We generated immortalized human LECs by lentiviral transduction of human telomerase (hTERT) and BMI-1 expression cassettes into primary LECs. Immortalized LECs showed an increased growth potential, reduced senescence, and elongated lifespan with maintenance of typical LEC morphology and marker expression for over 12 months while remaining nontransformed. Immortalized LECs were introduced in a microfluidic chip, comprising a free-standing extracellular matrix, where they formed a perfusable vessel-like structure against the extracellular matrix. A gradient of lymphangiogenic factors over the extracellular matrix gel induced the formation of luminated sprouts. Adding mouse colon cancer organoids adjacent to the lymphatic vessel resulted in a stable long-lived coculture model in which cancer cell-induced lymphangiogenesis and cancer cell motility can be investigated. Thus, the development of a stable immortalized lymphatic endothelial cell line in a membrane-free, perfused microfluidic chip yields a highly standardized lymphangiogenesis and lymphatic vessel-tumor cell coculture assay.
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Keywords: 3-lane OrganoPlate, BMI1, hTERT, LEC, Organ-on-a-chip, organ-on-a-chip, Endothelial Cells, Lymphatic Vessels, Coculture Techniques, Humans, Biology, Microfluidics, Biomedical Engineering, Biomaterials, Research Support, Non-U.S. Gov't, Journal Article
ISSN: 2373-9878
Publisher: American Chemical Society
Note: Funding Information: This work was supported by the Dutch Cancer Society/Koningin Wilhelmina Fonds (UU2014-6904, J.H.) and the Alexandre Suerman Young Talent Stipend (N.F.). Publisher Copyright: © 2021 American Chemical Society.
(Peer reviewed)