Compositions and methods of cell attachment

Patent metadata

Field	Value
Publication number	US-2018024117-A1
Application number	US-201715648293-A
Country	US
Kind code	A1
Filing date	Jul 12, 2017
Priority date	Jul 12, 2016
Publication date	Jan 25, 2018
Grant date	—

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.

First claim

We claim:

1 . A method of treating a microfluidic device, comprising: a) providing a microfluidic device comprising a microchannel comprising a surface, said microchannel in fluidic communication with a fluid source comprising fluid; b) covalently attaching one or more proteins or peptides to said microchannel surface so as to create a treated surface; and c) storing said microfluidic device.

2 . The method of claim 1 , wherein said storing is done at a controlled temperature below room temperature.

3 . The method of claim 1 , wherein said storing is done at between 2 and 10° C.

4 . The method of claim 1 , wherein said one or more covalently attached proteins is collagen I.

5 . The method of claim 4 , wherein said covalently attached collagen I is stored dry.

6 . The method of claim 1 , wherein said one or more covalently attached proteins is laminin.

7 . The method of claim 6 , wherein said covalently attached laminin is stored wet.

8 . The method of claim 1 , wherein said method further comprises: d) seeding viable cells on said treated surface so as to create attached cells; e) flowing fluid from said fluid source through said microchannel so as to create flowing conditions; and f) culturing said attached cells under said flow conditions such that said cells remain attached and viable for at least 14 days.

9 . The method of claim 8 , wherein said cells are hepatocytes.

10 . The method of claim 1 , wherein said microchannel surface is PDMS and is plasma treated prior to step b).

11 . A method of culturing cells, comprising: a) providing a microfluidic device comprising a microchannel comprising a surface, said microchannel in fluidic communication with a fluid source comprising fluid; b) covalently attaching a bifunctional crosslinker to said surface to create attached crosslinker, c) covalently attaching one or more proteins or peptides to said attached crosslinker as to create a treated surface; d) seeding viable cells on said treated surface so as to create attached cells; e) flowing fluid from said fluid source through said microchannel so as to create flowing conditions; and f) culturing said attached cells under said flow conditions such that said cells remain attached and viable for at least 7 days.

12 . The method of claim 11 , wherein said surface is a membrane and said membrane is micropatterned.

13 . The method of claim 11 , wherein said cells are skeletal muscle cells that align with said micropattern.

14 . The method of claim 11 , wherein said crosslinker is activated with UV light in the presence of a mask.

15 . A kit comprising:
a) a microfluidic device comprising a surface; b) a crosslinker comprising at least one light-reactive portion, and at least one chemically reactive portion; c) at least one protein or peptide; and d) a set of instructions.

16 . The kit of claim 15 , further comprising cells.

Assignees

• Emulate Inc

Inventors

• Levner Daniel
• Jang Kyung Jin
• Frase Jacob
• Kerns Jordan
• Varone Antonio
• Huh Dongeun

Classifications

• C12N2537/10

  Cross-linking · CPC title

• C12N5/0658

  Skeletal muscle cells, e.g. myocytes, myotubes, myoblasts · CPC title

• G01N33/5014

  for testing toxicity · CPC title

• C12N2533/30

  Synthetic polymers · CPC title

• G01N33/5032 Primary

  on intercellular interactions · CPC title