Modeling lymphangiogenesis in a three-dimensional culture system.
A lack of appropriate in vitro models of three-dimensional lymph vessel growth hampers the study of lymphangiogenesis. We developed a lymphatic ring assay--a potent, reproducible and quantifiable three-dimensional culture system for lymphatic endothelial cells that reproduces spreading of endothelial cells from a pre-existing vessel, cell proliferation, migration and differentiation into capillaries. In the assay, mouse thoracic duct fragments are embedded in a collagen gel, leading to the formation of lumen-containing lymphatic capillaries, which we assessed by electron microscopy and immunostaining. We developed a computerized method to quantify the lymphatic network. By applying this model to gene-deficient mice, we found evidence for involvement of the matrix metalloproteinase, MMP-2, in lymphangiogenesis. The lymphatic ring assay bridges the gap between two-dimensional in vitro models and in vivo models of lymphangiogenesis, can be used to exploit the potential of existing transgenic mouse models, and rapidly identify regulators of lymphangiogenesis.
Bruyère
Françoise
F
Laboratory of Tumor and Developmental Biology, Groupe Interdisciplinaire de Génoprotéomique Appliqué-Cancer, University of Liège, Avenue de l'Hôpital 3, B-4000 Liège, Belgium.
Melen-Lamalle
Laurence
L
Blacher
Silvia
S
Roland
Guy
G
Thiry
Marc
M
Moons
Lieve
L
Frankenne
Francis
F
Carmeliet
Peter
P
Alitalo
Kari
K
Libert
Claude
C
Sleeman
Jonathan P
JP
Foidart
Jean-Michel
JM
Noël
Agnès
A
D000818:Animals; D003198:Computer Simulation; D005260:Female; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D042583:Lymphangiogenesis / Q000187:drug effects / Q000502:physiology*; D008221:Lymphoid Tissue / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008954:Models, Biological; D010447:Peptide Hydrolases / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D058258:Serpin E2; D015843:Serpins / Q000235:genetics / Q000378:metabolism; D046509:Tissue Culture Techniques / Q000379:methods*
D036341:Intercellular Signaling Peptides and Proteins; D011480:Protease Inhibitors; D058258:Serpin E2; C511869:Serpine2 protein, mouse; D015843:Serpins; D010447:Peptide Hydrolases; D020778:Matrix Metalloproteinase 2; C522362:Mmp2 protein, mouse