Toshio Kumasaka1,2, Yoshito Hoshika3,2, Etsuko Kobayashi3,2, Keiko Mitani4,2, Makiko K Okura3,2, Young Kwon Hong5, Kazuhisa Takahashi3, Kuniaki Seyama3,2. 1. 1 Department of Pathology, Japanese Red Cross Medical Center , Tokyo, Japan . 2. 5 The Study Group of Pneumothorax and Cystic Lung Diseases , Tokyo, Japan . 3. 2 Division of Respiratory Medicine, Juntendo University Faculty of Medicine and Graduate School of Medicine , Tokyo, Japan . 4. 3 Division of Human Pathology, Juntendo University Faculty of Medicine and Graduate School of Medicine , Tokyo, Japan . 5. 4 Departments of Surgery and Biochemistry and Molecular Biology, Norris Comprehensive Cancer Center, University of Southern California , California.
Abstract
BACKGROUND: Lymphangioleiomyomatosis (LAM) is a tumor consisting of benign-looking neoplastic cells, but its wretched clinical outcome often resembles a malignant disease. LAM cell clusters (LCCs), unique microstructures commonly found in LAM-associated chylous effusion, are aggregates of LAM cells rimmed by lymphatic endothelium. LCCs seem to be crucial participants in the dissemination and progression of LAM. METHODS AND RESULTS: LCCs isolated from LAM-associated chylous effusion were embedded in a three-dimensional (3-D) culture gels, and then placed in a humidified CO2 incubator. During serial observations of their morphological changes, we found tube formations with lymphatic endothelial cells when LCCs settled side by side in 3-D gels. On the other hand, when LCCs were embedded separately enough to be isolated at their initial sites of settlement in the gels, each of LCCs gradually broke down, leaving a "cyst-like" hole after 7 days at the site where LCCs initially resided. Finally, we demonstrated that "cyst-like" hole formation in 3-D gels was inhibited with treatment with doxycycline or recombinant human VEGF receptor-3. CONCLUSIONS: We consider that our observation of this sequence in vitro illustrates how LCCs behave in vivo while enacting their important role in the progression of LAM. Our results indicate that the 3-D gel culture system for LCCs is a useful tool for exploring the effects of new therapeutic drugs under conditions when LCCs are constantly available.
BACKGROUND:Lymphangioleiomyomatosis (LAM) is a tumor consisting of benign-looking neoplastic cells, but its wretched clinical outcome often resembles a malignant disease. LAM cell clusters (LCCs), unique microstructures commonly found in LAM-associated chylous effusion, are aggregates of LAM cells rimmed by lymphatic endothelium. LCCs seem to be crucial participants in the dissemination and progression of LAM. METHODS AND RESULTS: LCCs isolated from LAM-associated chylous effusion were embedded in a three-dimensional (3-D) culture gels, and then placed in a humidified CO2 incubator. During serial observations of their morphological changes, we found tube formations with lymphatic endothelial cells when LCCs settled side by side in 3-D gels. On the other hand, when LCCs were embedded separately enough to be isolated at their initial sites of settlement in the gels, each of LCCs gradually broke down, leaving a "cyst-like" hole after 7 days at the site where LCCs initially resided. Finally, we demonstrated that "cyst-like" hole formation in 3-D gels was inhibited with treatment with doxycycline or recombinant human VEGF receptor-3. CONCLUSIONS: We consider that our observation of this sequence in vitro illustrates how LCCs behave in vivo while enacting their important role in the progression of LAM. Our results indicate that the 3-D gel culture system for LCCs is a useful tool for exploring the effects of new therapeutic drugs under conditions when LCCs are constantly available.
Authors: T Hayashi; M V Fleming; W G Stetler-Stevenson; L A Liotta; J Moss; V J Ferrans; W D Travis Journal: Hum Pathol Date: 1997-09 Impact factor: 3.466