| Autor: |
Frenkel N; UMC Utrecht Cancer Center, University Medical Center Utrecht and Utrecht University, Heidelberglaan 100, Utrecht 3584CX, The Netherlands., Poghosyan S; UMC Utrecht Cancer Center, University Medical Center Utrecht and Utrecht University, Heidelberglaan 100, Utrecht 3584CX, The Netherlands., Alarcón CR; UMC Utrecht Cancer Center, University Medical Center Utrecht and Utrecht University, Heidelberglaan 100, Utrecht 3584CX, The Netherlands., García SB; Mimetas BV, JH Oortweg 19, Leiden, The Netherlands., Queiroz K; Mimetas BV, JH Oortweg 19, Leiden, The Netherlands., van den Bent L; UMC Utrecht Cancer Center, University Medical Center Utrecht and Utrecht University, Heidelberglaan 100, Utrecht 3584CX, The Netherlands., Laoukili J; UMC Utrecht Cancer Center, University Medical Center Utrecht and Utrecht University, Heidelberglaan 100, Utrecht 3584CX, The Netherlands., Rinkes IB; UMC Utrecht Cancer Center, University Medical Center Utrecht and Utrecht University, Heidelberglaan 100, Utrecht 3584CX, The Netherlands., Vulto P; Mimetas BV, JH Oortweg 19, Leiden, The Netherlands., Kranenburg O; UMC Utrecht Cancer Center, University Medical Center Utrecht and Utrecht University, Heidelberglaan 100, Utrecht 3584CX, The Netherlands., Hagendoorn J; UMC Utrecht Cancer Center, University Medical Center Utrecht and Utrecht University, Heidelberglaan 100, Utrecht 3584CX, The Netherlands. |
| Abstrakt: |
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 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. |
