A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
Cell-derived matrices for studying cell proliferation and directional migration in a complex 3D microenvironment
Tekijät: Kaukonen R, Jacquemet G, Hamidi H, Ivaska J
Kustantaja: NATURE PUBLISHING GROUP
Julkaisuvuosi: 2017
Journal: Nature Protocols
Tietokannassa oleva lehden nimi: NATURE PROTOCOLS
Lehden akronyymi: NAT PROTOC
Vuosikerta: 12
Numero: 11
Aloitussivu: 2376
Lopetussivu: 2390
Sivujen määrä: 15
ISSN: 1754-2189
eISSN: 1750-2799
DOI: https://doi.org/10.1038/nprot.2017.107
Tiivistelmä
2D surfaces offer simple analysis of cells in culture, yet these often yield different cell morphologies and responses from those observed in vivo. Considerable effort has therefore been expended on the generation of more tissue-like environments for the study of cell behavior in vitro. Purified matrix proteins provide a 3D scaffold that better mimics the in vivo situation; however, these are far removed from the complex tissue composition seen in vivo. Cell-derived matrices (CDMs) offer a more physiologically relevant alternative for studying in vivo-like cell behavior in vitro. In the protocol described here, fibroblasts cultured on gelatin-coated surfaces are maintained in the presence of ascorbic acid to strengthen matrix deposition over 1-3 weeks. The resulting fibrillar CDMs are denuded of cells, and other cells are subsequently cultured on them, after which their behavior is monitored. We also demonstrate how to use CDMs as an in vivo-relevant reductionist model for studying tumor-stroma-induced changes in carcinoma cell proliferation and migration.
2D surfaces offer simple analysis of cells in culture, yet these often yield different cell morphologies and responses from those observed in vivo. Considerable effort has therefore been expended on the generation of more tissue-like environments for the study of cell behavior in vitro. Purified matrix proteins provide a 3D scaffold that better mimics the in vivo situation; however, these are far removed from the complex tissue composition seen in vivo. Cell-derived matrices (CDMs) offer a more physiologically relevant alternative for studying in vivo-like cell behavior in vitro. In the protocol described here, fibroblasts cultured on gelatin-coated surfaces are maintained in the presence of ascorbic acid to strengthen matrix deposition over 1-3 weeks. The resulting fibrillar CDMs are denuded of cells, and other cells are subsequently cultured on them, after which their behavior is monitored. We also demonstrate how to use CDMs as an in vivo-relevant reductionist model for studying tumor-stroma-induced changes in carcinoma cell proliferation and migration.
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