Device for culturing cells and method for making the same

Patent metadata

Field	Value
Publication number	US-2019241857-A1
Application number	US-201916261249-A
Country	US
Kind code	A1
Filing date	Jan 29, 2019
Priority date	Feb 5, 2018
Publication date	Aug 8, 2019
Grant date	—

Abstract

A device and method for cultivating cells are provided, wherein a cell cultivating layer is formed on a surface of a substrate, and a temperature-responsive layer having a plurality of temperature-responsive polymer with magnetic objects is formed on a surface of the cell cultivating layer. When a controlling characteristic exerted on the temperature-responsive layer is varied, a plurality of cells can be adhered on the cell cultivating layer or detached therefrom. When the device is utilized, a temperature control step is operated to control environmental temperature at a first temperature for inducing temperature-responsive polymers becoming hydrophobic whereby the plurality of cells are adhered on the cell cultivating layer. In addition, the alternating magnetic field is utilized to rise temperature of the temperature-responsive polymers so that the plurality of cells can be kept being adhered on the cell cultivating layer at a second temperature lower than the first temperature.

First claim

What is claimed is:

1 . A device for cultivating cells, comprising:
a substrate; a cell cultivating layer, formed on a surface of the substrate; and a temperature-responsive layer, formed on a surface of the cell cultivating layer, wherein the temperature-responsive layer comprises a plurality of magnetic temperature-responsive polymers, each of which comprises a temperature-responsive polymer and a plurality of magnetic objects; wherein a plurality of cells are attached to the cell cultivating layer or detached from the cell cultivating layer according to a variation of at least one controlling characteristic.

2 . The device of claim 1 , wherein the cell cultivating layer comprises polyethylenimine, and the temperature-responsive polymer is (poly(N-Isopropylacrylamide)).

3 . The device of claim 1 , wherein the controlling characteristic is physical characteristic or a chemical characteristic, wherein the physical characteristic is temperature, magnetic field, or the combination of temperature and magnetic field, and the chemical characteristic is PH value, ionic condition, isoelectric point, or material compound.

4 . The device of claim 1 , further comprising a magnetic structure having a structural pattern, wherein the magnetic structure is arranged between the cell cultivating layer and the substrate or is arranged at external side of the substrate.

5 . The device of claim 4 , wherein the structural pattern comprises a plurality of concentric geometric patterns.

6 . The device of claim 5 , wherein the plurality of magnetic temperature-responsive polymers are arranged on at least one specific position of the structural pattern or are arranged along the structural pattern.

7 . The device of claim 6 , wherein the plurality of cells are attracted by a dissipative field around corners of the structural pattern, so that the cells are concentratedly attached to portions around corners where the magnetic temperature-responsive polymers are formed thereon so as to form a patterning cell cultivating substrate.

8 . A method for forming a cell cultivating device, comprising steps of:
forming a plurality of magnetic temperature-responsive polymers, each of which comprises a first temperature-responsive polymer having a plurality of magnetic objects; providing a substrate; forming a cell cultivating layer on the substrate; and forming a temperature-responsive layer on the cell cultivating layer by utilizing the plurality of magnetic temperature-responsive polymers thereby forming the cell cultivating device; wherein a plurality of cells are attached to the cell cultivating layer or detached from the cell cultivating layer according to a variation of at least one controlling characteristic.

9 . The method of claim 8 , wherein before forming the cell cultivating layer on the substrate, it further comprises a step of forming a magnetic film layer on the substrate, wherein the magnetic film layer further comprises a structural pattern.

10 . The method of claim 9 , wherein the plurality of magnetic temperature-responsive polymers are arranged on at least one specific position of the structural pattern after forming the temperature-responsive layer.

11 . The method of claim 9 , wherein the temperature-responsive layer is formed by coating a mixed liquid comprising a plurality of second temperature-responsive polymers mixed with the magnetic temperature-responsive polymers on the cell cultivating layer wherein area corresponding to the structural pattern has the magnetic temperature-responsive polymers formed thereon while the second temperature-responsive polymers are formed on the cell cultivating layer without having the structural pattern.

12 . The method of claim 8 , further comprising a step of arranging a permanent magnet having a structural pattern at one side of the substrate.

13 . The method of claim 11 , wherein the plurality of magnetic temperature-responsive polymers are arranged on at least one specific position of the structural pattern or are arranged along the structural pattern.

14 . The method of claim 12 , wherein the plurality of cells are attracted by a dissipative field around corners of the structural pattern, so that the cells are concentratedly attached to portions around corners where the magnetic temperature-responsive polymers are formed thereon so as to form a patterning cell cultivating substrate.

15 . The method of claim 10 , wherein the controlling characteristic is physical characteristic combining temperature and magnetic field, and the magnetic temperature-responsive polymers are contracted under a first temperature whereby the cells are attached on the cell cultivating layer, while the magnetic temperature-responsive polymers corresponding to the specific position of the structural pattern maintaining contracting status under a second temperature with a variation of the magnetic field whereby the cells located at the specific position are attached by the cell cultivating layer.

16 . A method for cell cultivation, comprising steps of:
providing a cell cultivating device comprising a substrate, a cell cultivating layer, formed on a surface of the substrate, a temperature-responsive layer, formed on a surface of the cell cultivating layer, wherein the temperature-responsive layer comprises a plurality of magnetic temperature-responsive polymers, each of which comprises a first temperature-responsive polymer and a plurality of magnetic objects, and a magnetic structure having a structural pattern; enabling a plurality of cells to be attached on the cell cultivating device at a first working temperature; providing an alternative magnetic field; and lowering the first working temperature to a second working temperature so as to form patterning cell cultivating substrate.

17 . The method of claim 18 , wherein the temperature-responsive layer is formed by coating a mixed liquid comprising a plurality of second temperature-responsive polymers mixed with the magnetic temperature-responsive polymers on the cell cultivating layer wherein area corresponding to the structural pattern has the magnetic temperature-responsive polymers formed thereon while the second temperature-responsive polymers are formed on the cell cultivating layer without having the structural pattern.

18 . The method of claim 16 , wherein the plurality of cells are attracted by a dissipative field around corners of the structural pattern, so that the cells are concentratedly attached to portions around corners where the magnetic temperature-responsive polymers are formed thereon so as to form the patterning cell cultivating substrate.

19 . The method of claim 16 , wherein the magnetic structure is arranged between the cell cultivating layer and the substrate or is arranged at external side of the substrate.

20 . The method of claim 16 , wherein the structural pattern comprises a plurality of concentric geometric patterns.

Assignees

• Univ Chung Yuan Christian

Inventors

• Ger Tzong-Rong
• Chen Tan-Yueh
• Su Chao-Ming
• Huang Yin-Chou
• Dong Fan
• Kuo Yu-Chi

Classifications

• C12N2535/10

  Patterned coating · CPC title

• C12N2523/00

  Culture process characterised by temperature · CPC title

• C12N2533/30

  Synthetic polymers · CPC title

• C12N2527/00

  Culture process characterised by the use of mechanical forces, e.g. strain, vibration · CPC title

• C12M41/12

  of temperature (controlling the temperature of chemical or physical processes B01J19/0013, heating or cooling apparatus for laboratory use B01L7/00) · CPC title