Cell isolation method and cell trapping filter

Patent metadata

Field	Value
Publication number	US-10538732-B2
Application number	US-201515523237-A
Country	US
Kind code	B2
Filing date	Oct 29, 2015
Priority date	Oct 31, 2014
Publication date	Jan 21, 2020
Grant date	Jan 21, 2020

Abstract

A cell isolation method includes: a cell trapping step of allowing a test liquid to pass through a cell trapping filter which has a plurality of through-holes in the thickness direction, thereby trapping isolation target cells contained in the test liquid on one surface of the cell trapping filter; a gel embedding step of introducing a stimulus-responsive hydrogel onto the one surface of the cell trapping filter on which the cells have been trapped in the cell trapping step, thereby embedding the cells in the stimulus-responsive hydrogel; a gel hardening step of applying a stimulus to the stimulus-responsive hydrogel in which the cells are embedded, thereby hardening the stimulus-responsive hydrogel; and a detachment step of detaching the stimulus-responsive hydrogel that was hardened in the gel hardening step from the cell trapping filter.

First claim

The invention claimed is:

1. A cell isolation method comprising:
a cell trapping step of allowing a test liquid to pass through a cell trapping filter which has a plurality of through-holes in the thickness direction, thereby trapping isolation target cells contained in the test liquid on one surface of the cell trapping filter;
a gel embedding step of introducing a stimulus-responsive hydrogel onto the one surface of the cell trapping filter on which the cells have been trapped in the cell trapping step, thereby embedding the cells in the stimulus-responsive hydrogel;
a gel hardening step of applying a stimulus to the stimulus-responsive hydrogel in which the cells are embedded, thereby hardening the stimulus-responsive hydrogel; and
a detachment step of detaching the stimulus-responsive hydrogel that was hardened in the gel hardening step from the cell trapping filter.

2. The cell isolation method according to claim 1 , wherein only the periphery of the isolation target cells in the stimulus-responsive hydrogel in which the cells are embedded is locally hardened in the gel hardening step.

3. The method according to claim 1 , wherein the distance between the adjacent through-holes on the side of the one surface is 80 μm or more.

4. The method according to claim 2 , wherein the distance between the adjacent through-holes on the side of the one surface is 80 μm or more.

5. The cell isolation method according to claim 1 , wherein the stimulus-responsive hydrogel comprises a material selected from the group consisting of polyethyleneglycol diacrylate, poly(N-isopropylacrylamide) and chitosan gel.

6. The cell isolation method according to claim 1 , wherein the stimulus-responsive hydrogel is photocurable.

7. The cell isolation method according to claim 1 , wherein the stimulus used in the gel hardening step is light from a light source.

8. The cell isolation method according to claim 1 , wherein the stimulus used in the gel hardening step is ultraviolet laser light.

9. The cell isolation method according to claim 1 , wherein a height of the stimulus-responsive hydrogel formed on the cell trapping filter in the gel embedding step is from 100 μm to 600 μm.

10. The cell isolation method according to claim 1 , wherein a height of the stimulus-responsive hydrogel introduced formed on the cell trapping filter in the gel embedding step is from 200 μm to 300 μm.

11. The cell isolation method according to claim 1 , wherein a width of the stimulus-responsive hydrogel after the gel hardening step is from 100 μm to 400 μm.

12. The cell isolation method according to claim 1 , wherein a height of the stimulus-responsive hydrogel after the gel hardening step is from 100 μm to 500 μm.

13. The cell isolation method according to claim 1 , wherein a shape of an opening of the through-holed of the cell trapping filter is a round, an oval, a square, a rectangular, a rectangle with rounded corners, or a polygon.

14. The cell isolation method according to claim 1 , wherein the stimulus-responsive hydrogel after the gel hardening step has a shape in which a central part of the hydrogel has a line symmetry narrow.

Assignees

• National Univ Corporation Tokyo Univ Of Agriculture And Technology

Inventors

• Yoshino Tomoko
• Tanaka Tsuyoshi
• Matsunaga Tadashi
• NEGISHI Ryo
• Kanbara Hisashige
• Nakamura Seita

Classifications

• G01N33/575

  for cancer · CPC title

• C12M3/06

  with filtration, ultrafiltration, inverse osmosis or dialysis means · CPC title

• C12M1/12

  with sterilisation, filtration or dialysis means · CPC title

• C12M1/28

  being part of container · CPC title

• C12M33/14 Primary

  with filters, sieves or membranes · CPC title