Cryopreservation of biological cells and tissues

• US 9,700,038 B2
• Filed: 09/22/2014
• Issued: 07/11/2017
• Est. Priority Date: 02/25/2009
• Status: Active Grant

• Current Assignee: Genea Ip Holdings Pty Limited
• Original Assignee: Genea Limited
• Inventors: Stojanov, Tomas
• Primary Examiner(s): Kim, Taeyoon
• Application Number:
  US14/492,477
• Publication Number:
  US 20150011000A1
• Time in Patent Office: 1,023 Days
• CPC Class Codes: A01N 1/0257 Stationary or portable vessels generating cryogenic temperatures
• CPC Class Codes: A01N 1/0268 Carriers for immersion in cryogenic fluid, both for slow-freezing and vitrification, e.g. open or closed "straws" for embryos, oocytes or semen

Abstract

The method involves placing an oocyte cell in a cell holder (1), securing the cell holder in a treatment station, applying a treatment solution to the cell by washing the cell with the solution, and rapidly cooling the cell holder and cell to a predetermined cryopreservation temperature for cryopreservation of the cell. The cell is cooled at a high rate sufficient to permit vitrification of the cell and any surrounding treatment solution to occur. The cell is then maintained at or below a predetermined storage temperature for storage. The method allows multiple cells to be treated simultaneously each secured within a respective cell holder.

Claims

1. An organic cell cryopreservation apparatus for use in the vitrification of at least one organic cell, said apparatus including:

a plurality of cell holders, each having a filter to retain one of the at least one organic cell while allowing for fluid to pass from the cell holder;

a securing frame for securing each cell holder in a predetermined orientation, the securing frame comprising;

an array of cell holder receiving portions, each configured to receive one of the cell holders; and

at least one securing formation for securing the cell holders in the cell holder receiving portions; and

a treatment station for washing said at least one organic cell, said treatment station including;

a temperature controller;

at least one pipette to dispense a predetermined measure of treatment solution into each cell holder; and

a docking formation for engaging a corresponding docking formation on the securing frame thereby aligning said securing frame into a predetermined orientation with said treatment station.

2. An apparatus according to claim 1, said apparatus further including a cooling station for cooling said cells to a cryopreservation temperature.

3. An apparatus according to claim 2, wherein said cooling station includes a liquid nitrogen cooled block.

4. An apparatus according to claim 2, wherein the treatment station comprises the cooling station.

5. An apparatus according to claim 1, the treatment station further comprising a drain for receiving treatment fluid draining from said cell holder.

6. An apparatus according to claim 1, said frame having a drainage path for directing treatment fluid passing through said filtration arrangement and from said cell holder to a drainage outlet.

7. An apparatus according to claim 1, wherein said filtration arrangement includes at least one of:

a mesh;

a membrane;

a fibrous material; and

a perforated section.

8. An apparatus according to claim 1, wherein the cell-holder comprises a cup-shaped holding vessel.

9. An apparatus according to claim 8, wherein the cell-holder comprises converging sidewalls.

10. An apparatus according to claim 1, wherein the treatment station comprises a multi-headed pipette to simultaneously dispense predetermined measures of treatment solution into the plurality of cell holders.

18. An apparatus according to claim 1, wherein the treatment station is automated to dispense predetermined measures of treatment solution into the cell holders secured in the securing frame and drain at least part of the treatment solution away from the cell holders.

19. An apparatus according to claim 1, wherein the treatment station further comprises a temperature controller.

20. An apparatus according to claim 2, wherein liquid nitrogen is used in the cooling station for cooling said cells to a cryopreservation temperature.

11. An apparatus for vitrification of organic cells, the apparatus comprising:

a plurality of cell holders, each cell holder comprising a cup-shaped vessel with converging sidewalls and a filter for retaining an organic cell in the cell holder while allowing for fluid to pass from the cell holder;

a securing frame defining an array of cell holder receiving portions and comprising at least one securing formation for securing the cell holders in the cell holder receiving portions; and

a treatment station comprising a docking formation configured to hold the securing frame in a predetermined orientation relative to the treatment station, and at least one pipette for dispensing a predetermined measure of treatment solution into each cell holder.

12. An apparatus according to claim 11, wherein the treatment station comprises a multi-headed pipette to simultaneously dispense predetermined measures of treatment solution into each of the plurality of cell holders.

13. An apparatus according to claim 11, said apparatus further including a cooling station for cooling said cells to a cryopreservation temperature.

14. An apparatus according to claim 13, wherein said cooling station includes a liquid nitrogen cooled block.

15. An apparatus according to claim 13, wherein the treatment station comprises the cooling station.

16. An apparatus according to claim 11, wherein the treatment station is automated to dispense predetermined measures of treatment solution into the cell holders secured in the securing frame and drain at least part of the treatment solution away from the cell holders.

17. An apparatus according to claim 11, wherein the treatment station further comprises a temperature controller.

21. An apparatus according to claim 13, wherein liquid nitrogen is used in the cooling station for cooling said cells to a cryopreservation temperature.