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Development of a cultured lymphatic muscle cell line from rat cervical duct

All of the functions of the lymphatic systems rely on a regulated lymph flow. Lymphatic muscle is responsible for the phasic and tonic contractions used to generate and regulate lymph flow. Yet our understanding of the mechanisms regulating lymphatic muscle contraction is poor. To study the regulation of lymphatic contractile function suitable cultured lymphatic muscle cell lines would be extremely useful. We have developed a technique to isolate and culture the muscle cells from known, single lymphatics. We have previously used this technique to develop cultured lymphatic muscle lines from rat mesenteric microlymphatics. To compare these cells to muscle cells from large conduit lymphatics, we established cultured muscle cell lines from rat cervical duct. These vessels were ~300–400 μm in diameter. To isolate muscle cells from these vessels, a suitable cervical duct was located and carefully dissected from the surrounding tissue. The vessel was carefully cleaned and placed on a gelatin coated plastic culture dish. The vessel was incubated until small patches of cells are seen growing off the vessel 3–4 days later. The explanted vessel was then removed. The remaining cells were cultured, passed and screened for smooth muscle phenotype by morphology and staining for alpha‐vascular smooth muscle actin. The defined cervical duct muscle cells were then passed and used in studies. (NIH HL‐075199)