title: Isolation of human lymphatic malformation endothelial cells, their in vitro characterization and in vivo survival in a mouse xenograft model.
abstract: Human lymphatic vascular malformations (LMs), also known as cystic hygromas or lymphangioma, consist of multiple lymphatic endothelial cell-lined lymph-containing cysts. No animal model of this disease exists. To develop a mouse xenograft model of human LM, CD34(Neg)CD31(Pos) LM lymphatic endothelial cells (LM-LEC) were isolated from surgical specimens and compared to foreskin CD34(Neg)CD31(Pos) lymphatic endothelial cells (LECs). Cells were implanted into a mouse tissue engineering model for 1, 2 and 4 weeks. In vitro LM-LECs showed increased proliferation and survival under starvation conditions (P < 0.0005 at 48 h, two-way ANOVA), increased migration (P < 0.001, two-way ANOVA) and formed fewer (P = 0.029, independent samples t test), shorter tubes (P = 0.029, independent samples t test) than foreskin LECs. In vivo LM-LECs implanted into a Matrigel™-containing mouse chamber model assembled to develop vessels with dilated cystic lumens lined with flat endothelium, morphology similar to that of clinical LMs. Human foreskin LECs failed to survive implantation. In LM-LEC implanted chambers the percent volume of podoplanin(Pos) vessels was 1.18 ± 2.24 % at 1 week, 6.34 ± 2.68 % at 2 weeks and increasing to 7.67 ± 3.60 % at 4 weeks. In conclusion, the significantly increased proliferation, migration, resistance to apoptosis and decreased tubulogenesis of LM-LECs observed in vitro is likely to account for their survival and assembly into stable LM-like structures when implanted into a mouse vascularised chamber model. This in vivo xenograft model will provide the basis of future studies of LM biology and testing of potential pharmacological interventions for patients with lymphatic malformations.
authors.affiliation: O'Brien Institute, 42 Fitzroy Street, Fitzroy, VIC, 3065, Australia, zerina.lokmic@mcri.edu.au.
authors.forename: Zerina
authors.initials: Z
authors.lastname: Lokmic
authors.forename: Geraldine M
authors.initials: GM
authors.lastname: Mitchell
authors.forename: Nicholas
authors.initials: N
authors.lastname: Koh Wee Chong
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Bastiaanse
authors.forename: Yi-Wen
authors.initials: YW
authors.lastname: Gerrand
authors.forename: Yiping
authors.initials: Y
authors.lastname: Zeng
authors.forename: Elizabeth D
authors.initials: ED
authors.lastname: Williams
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Penington
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D049109:Cell Proliferation; D002469:Cell Separation; D002470:Cell Survival; D002648:Child; D002675:Child, Preschool; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology / Q000637:transplantation; D005260:Female; D006085:Graft Survival; D064593:Heterografts; D006801:Humans; D007223:Infant; D042601:Lymphatic Vessels / Q000002:abnormalities / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D016513:Mice, SCID; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods
chemical_list: D018952:Antigens, CD34; D019408:Platelet Endothelial Cell Adhesion Molecule-1