Immortalized Mouse Dermal Lymphatic Endothelial Cells-SV40

Immortalized mouse dermal lymphatic endothelial cells (LECs) are generated by transducing primary murine dermal LECs with a retroviral or lentiviral vector encoding the simian virus 40 (SV40) large T antigen. The distinct advantages of this cell line are threefold. First, indefinite proliferation and scalability – SV40‑immortalised LECs can be propagated beyond 30 passages with consistent doubling times, ensuring a reliable and abundant supply for long‑term and high‑throughput studies, eliminating the batch‑to‑batch variability inherent to primary isolates. Second, faithful retention of lymphatic identity – these cells maintain characteristic cobblestone morphology and express the full panel of definitive lymphatic markers, including podoplanin, Prox‑1 (a master transcriptional regulator), VEGFR‑3 (Flt‑4), and LYVE‑1, while remaining negative for blood vascular markers such as CD34. This molecular signature is stably maintained over extended culture, confirming their specialized lineage. Third, functional competence – immortalized LECs form capillary‑like tubular networks on Matrigel, actively migrate in response to VEGF‑C and sphingosine‑1‑phosphate (S1P), and upregulate ICAM‑1 and VCAM‑1 upon TNF‑α stimulation, faithfully recapitulating the inflammatory activation seen in vivo. Their ability to support lymphocyte adhesion and transmigration makes them an invaluable tool for studying immune surveillance and metastatic dissemination. Notably, they remain non‑tumorigenic and anchorage‑dependent, ensuring phenotypic stability without malignant transformation. Together, SV40‑immortalised mouse dermal LECs offer a robust, phenotypically authentic, and experimentally tractable platform that bridges fundamental lymphatic biology with translational research in cancer, immunology, and cardiovascular disease.