System and method for fusing cells

Patent metadata

Field	Value
Publication number	US-9249385-B1
Application number	US-201414574635-A
Country	US
Kind code	B1
Filing date	Dec 18, 2014
Priority date	Dec 18, 2014
Publication date	Feb 2, 2016
Grant date	Feb 2, 2016

Abstract

The present invention is concerned with a system for fusion at least two cells. The system has an optical tweezers system for generating moving optical traps for manipulating the cells, an optical scissors system for cutting cell membrane of the cells and inducing fusion of the cells, an incubation system for providing cell culture in which the cells suspend and cell culture environment for the cells, and a visual detection system allowing visual monitoring of the cells undergoing fusion.

First claim

The invention claimed is:

1. A system for fusion at least two cells comprising:—
a) an optical tweezers system for generation of a plurality of optical traps for manipulating the cells in isolation;
b) an optical scissor system for cutting cell membrane of the cells and inducing fusion of the cells;
c) an incubation system for providing cell culture in which the cells suspend and cell culture environment for the cells;
d) a visual detection system allowing visual monitoring of the cells undergoing fusion; and
e) a control system for controlling motion and position of a stage supporting the cells, controlling operation diffraction optical elements, and cutting frequency and time of optical scissors.

2. A system as claimed in claim 1 , wherein said system is free from using chemical factors, viral factors or antibiotics factors, in inducing the fusion of the cells, or free from means to effect electro-fusion.

3. A system as claimed in claim 2 , wherein said optical tweezers system is a holographic optical tweezers system.

4. A system as claimed in claim 2 , wherein said optical tweezers system is configured to produce a continuous-wave laser source.

5. A system as claimed in claim 2 , comprising means for splitting and steering single light beam into multiple beams.

6. A system as claim in claim 5 , wherein said means includes diffractive laser optical elements.

7. A system as claimed in claim 2 , wherein said optical scissor system is a pulsed UV laser system.

8. A system as claimed in claim 7 , comprising a telescope for expanding pulsed laser beam from said pulsed UV laser system.

9. A system as claimed in claim 2 , wherein said incubation system is a stage top incubation system including a stage top incubator provided with environment controls.

10. A system as claimed in claim 5 , wherein said environment controls include temperature control and carbon dioxide (CO 2 ) concentration control.

11. A system as claimed in claim 1 , wherein said vision detection system includes a microscope provided with a motorized stage, a light source and CCD camera.

12. A system as claimed in claim 11 , wherein said vision detection system is configured for bright-field imaging.

13. A system as claimed in claim 11 , wherein said vision detection system is configured for fluorescence imaging.

14. A system as claimed in claim 8 , wherein the UV laser system is adapted to deliver pulsed UV laser of substantially 355 nm in wavelength.

15. A system as claimed in claim 1 , wherein said control system is configured to control generation of one or more optical traps by said diffraction optical elements and movement of said one or more optical traps.

16. A system as claimed in claim 1 , comprising a container for containing the cells, bottom of said container being transparent at wavelength of lasers from said optical systems.

17. A method of using a system as claimed in claim 1 for fusing at least two cells together, comprising:—
a) manipulating the cells by using optical tweezers to form a cell pair or a cell chain shape; and
b) fusing cell membrane of the cells by subjecting the cell membrane with pulsed UV laser.

18. A method for fusing at least two cells together, comprising:—
a) manipulation of the cells by using optical tweezers to form a cell pair or a cell chain; and
b) fusion of cell membrane of the cells by subjecting the cell membrane with pulsed UV laser;
wherein said manipulation includes controlling movement of motorized stage supporting the cells and position of said optical tweezers.

19. A method as claimed in claim 18 , comprising generation of an optical trap for moving the cells close together or in contact with each other for forming the cell pair or cell chain.

20. A method as claimed in claim 18 , wherein the cells fused together is a viable bi-nucleate or multi-nucleate cell.

21. A method as claimed in claim 18 , wherein the cells before fusion are in condition of suspended cells or adherent cells.

22. A method as claimed in claim 18 , wherein the cells include a stem cell and a somatic cell.

23. A method as claimed in claim 17 , comprising generation of an optical trap for moving the cells close together or in contact with each other for forming the cell pair or cell chain.

24. A method as claimed in claim 17 , wherein the cells fused together is a viable bi-nucleate or multi-nucleate cell.

25. A method as claimed in claim 17 , wherein the cells before fusion are in condition of suspended cells or adherent cells.

26. A method as claimed in claim 17 , wherein the cells include a stem cell and a somatic cell.

Assignees

• Univ City Hong Kong

Inventors

• Sun Dong
• Chen Shuxun
• Wang Ran

Classifications

• C12M35/02 Primary

  Electrical or electromagnetic means, e.g. for electroporation or for cell fusion · CPC title

• C12N15/02

  Preparation of hybrid cells by fusion of two or more cells, e.g. protoplast fusion {(monoclonal antibodies C07K16/00; apparatus for cell fusion C12M)} · CPC title