Research Project: Engineering biomimetic niches to study tumorigenesis and stages of tumor progression
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Contributors
Funders
ID
TB.00478
Authors
Öztürk, Ece
Faculty Member
Publications
Growth and organotypic branching of lung-specific microvascular cells on 2D and in 3D lung-derived matrices
(N/A, 2024) Özkan, Sena Nur; Öztürk, Ece; School of Medicine; Graduate School of Health Sciences; Yes; GRADUATE SCHOOL OF HEALTH SCIENCES; SCHOOL OF MEDICINE
Tissue-specific endothelial cells have vital roles in maintenance and functioning of native tissues with constant reciprocal crosstalk with resident cells. Three-dimensional (3D) physio-mimetic in vitro models which incorporate lung-specific microvasculature are needed to model lung-related diseases which involve modulation of endothelial cell behavior like cancer. In this study, we investigated the growth kinetics, morphological changes and responses to biological cues of lung microvasculature on two-dimensional (2D) and in lung matrix-derived 3D hydrogels. HUVEC and HULEC-5a cells were cultured on 2D and compared for their growth, morphologies, and responses to varying growth medium formulations. Brightfield and immunofluorescence imaging was performed to assess differences in morphology. For 3D cultures, native bovine lungs were decellularized, lyophilized, solubilized, and reconstituted into hydrogel form in which endothelial cells were embedded. Cell growth and organotypic branching was monitored in 3D hydrogels in the presence of varying biological cues including lung cancer cell secretome. HUVEC and HULEC-5a cells demonstrated comparable growth and morphology on 2D. However, in 3D lung-derived ECM hydrogels, tissue-specific HULEC-5a cells exhibited much better adaptation to their microenvironment, characterized by enhanced organotypic branching and longer branches. HULEC-5a growth was responsive to lung cancer cell-conditioned medium in both 2D and 3D conditions. In 3D, the concentration of ECM ligand significantly affected cell growth in long-term culture where molecular crowding had an inhibitory role. Our data reveals that HULEC-5a cells offer a reliable alternative to frequently pursued HUVECs with comparable growth and morphology. Due to their intrinsic program for cellular crosstalk with resident cells, the use of tissue-specific endothelium constitutes a vital aspect for modeling physiological and pathological processes. Furthermore, our study is the first demonstration of the synergy between lung-specific microvasculature with lung-specific ECM within a 3D in vitro model.
Different decellularization methods in bovine lung tissue reveals distinct biochemical composition, stiffness, and viscoelasticity in reconstituted hydrogels
(American Chemical Society (ACS), 2023) Bulutay, Pınar; Fırat, Pınar Arıkan; Karaoğlu, İsmail Can; Kızılel, Seda; Kuşoğlu, Alican; Örnek, Deniz; Özkan, Sena Nur; Sarıca, Sevgi; Solcan, Nuriye; Yangın, Kardelen; Department of Chemical and Biological Engineering; Graduate School of Health Sciences; Graduate School of Sciences and Engineering; KUTTAM (Koç University Research Center for Translational Medicine); School of Medicine; Yes; College of Engineering; GRADUATE SCHOOL OF HEALTH SCIENCES; GRADUATE SCHOOL OF SCIENCES AND ENGINEERING; Research Center; SCHOOL OF MEDICINE
Extracellula r matri x (ECM)-derived hydrogels are in demand for use in lung tissue engineering to mimic the native microenvironment of cells in vitro. Decellularization of native tissues has been pursued for preser v i n g organotypic ECM while eliminating cellular content and reconstitution into scaffolds which allows re-cellularization for modeling homeostasis, regeneration, or diseases. Achieving mechanical stabi l i t y and understanding the effects of the decellularization process on mechanical parameters of the reconstituted ECM hydrogels present a challenge in the field. Stiffness and viscoelasticity are important characteristics of tissue mechanics that regulate crucial cellular processes and their in vitro representation in engineered models is a current aspiration. The effect of decellulariza-tion on viscoelastic properties of resulting ECM hydrogels has not yet been addressed. The aim of this study was to establish bovine lung tissue decellularization for the first time via pursuing four different protocols and characterization of reconstituted decellularized lung ECM hydrogels for biochemical and mechanical properties. Our data reveal that bovine lungs provide a reproducible alternative to human lungs for disease modeling with optimal retention of ECM components upon decellularization. We demonstrate that the decellularization method significa n t l y affects ECM content, stiffness, and viscoelastic properties of resulting hydrogels. Lastly, we examined the impact of these aspects on viabi l i t y , morphology, and growth of lung cancer cells, healthy bronchial epithelial cells, and patient-derived lung organoids.
