Compositions and methods of cell attachment
AssigneeEmulate, Inc.
InventorsDaniel Levner, Kyung Jin Jang, Jacob Fraser, S. Jordan Kerns, Antonio Varone, Dongeun Huh
Life of this patent
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Abstract
Compositions, devices and methods are described for improving adhesion, attachment, and/or differentiation of cells in a microfluidic device or chip. In one embodiment, one or more ECM proteins are covalently coupled to the surface of a microchannel of a microfluidic device. The microfluidic devices can be stored or used immediately for culture and/or support of living cells such as mammalian cells, and/or for simulating a function of a tissue, e.g., a liver tissue, muscle tissue, etc. Extended adhesion and viability with sustained function over time is observed.
Claims10
A method of culturing cells, comprising:
- a) providing a microfluidic device comprising a microchannel comprising a polydimethylsiloxane surface, said polydimethylsiloxane surface reacted with an ultraviolet light reactive portion of a N-sulfosuccinimidyl-6-(4′-azido-2′-nitrophenylamino) hexanoate heterobifunctional crosslinker using an ultraviolet light source, said microchannel in fluidic communication with a fluid source comprising a fluid;
- b) covalently attaching one or more extracellular matrix proteins to a chemically-reactive portion of said heterobifunctional crosslinker so as to create a treated surface;
- c) seeding viable epithelial cells on said treated surface so as to create attached epithelial cells;
- d) flowing said fluid from said fluid source through said microchannel so as to create flowing conditions;
- and
- e) culturing said attached epithelial cells under said flow conditions such that said attached epithelial cells remain attached and viable for at least 7 days.
A method of culturing cells, comprising:
- a) providing a microfluidic device comprising a microchannel comprising a polydimethylsiloxane surface, said surface reacted with an ultraviolet light reactive portion of a N-sulfosuccinimidyl-6-(4′-azido-2′-nitrophenylamino) hexanoate crosslinker using a source of ultraviolet light, said microchannel in fluidic communication with a fluid source comprising a fluid;
- b) covalently attaching one or more extracellular matrix proteins to a chemically-reactive portion of said crosslinker so as to create a treated surface;
- c) seeding viable hepatocytes on said treated surface so as to create attached cells;
- d) flowing said fluid from said fluid source through said microchannel so as to create flowing conditions;
- and
- e) culturing said attached cells under said flow conditions such that said cells remain attached and viable for at least 7 days.
Dependencies are inferred from the claim text.