Product and method for cell preservation

Patent metadata

Field	Value
Publication number	US-10959423-B2
Application number	US-201515125871-A
Country	US
Kind code	B2
Filing date	Feb 19, 2015
Priority date	Mar 31, 2014
Publication date	Mar 30, 2021
Grant date	Mar 30, 2021

Abstract

The present invention relates to a product and method for cell preservation, such as cell banking of any type of cells for subsequent scientific or medical use. More closely, the invention relates to a product and a method for cryopreservation of cells using vacuum tubes provided with freeze media/cryopreservation liquid.

First claim

The invention claimed is:

1. A method for cryopreservation of cells from a bioreactor, the method comprising,
cultivating the cells in the bioreactor as a perfusion culture to 10-200 MVC/mL,
providing an access device having a needleless connector configured for forming a needleless connection to the bioreactor;
using the needleless connector of the access device to form a needleless connection with the bioreactor containing the cells to access the cells;
sampling or harvesting the cells from the device into a vacuum tube provided with freeze media by connecting the vacuum tube with the access device such that the cells are instantly mixed with the freeze media; and
freezing said vacuum tube for cryopreservation of the cells.

2. The method according to claim 1 , further comprising adding a freeze media into an empty vacuum tube to form the vacuum tube provided with freeze media.

3. The method according to claim 1 , wherein the freeze media is DMSO, glycerol or methyl cellulose.

4. The method according to claim 1 , wherein the vacuum tube is filled with 5-20% v/v of freeze media.

5. The method according to claim 1 , wherein one or more steps of the method are automated.

6. The method according to claim 1 , further comprising incubating the cells mixed with the freeze media in the vacuum tube before freezing.

7. The method according to claim 1 , wherein the access device comprises a needle for inserting into the vacuum tube to form the connection between the vacuum tube and the access device.

8. A method for direct cryopreservation of cultivated cells from a bioreactor, the method comprising,
cultivating the cells in the bioreactor as a perfusion culture to 10-200 MVC/mL,
providing an access device having a connector configured for forming a needleless connection to a bioreactor and a sleeved needle configured for insertion into a vacuum tube, wherein the connector and the needle are in fluid communication with each other;
using the connector of the access device to form a needleless connection with the bioreactor to access the cultivated cells;
sampling or harvesting the cells from the bioreactor by inserting a vacuum tube provided with 5-20% v/v of DMSO into the needle of the access device such that the cells are instantly mixed with the DMSO; and
freezing said vacuum tube for cryopreservation of the cells,
wherein one or more steps of the method are automated.

9. The method according to claim 8 , further comprising adding DMSO into an empty vacuum tube to form the vacuum tube provided with 5-20% v/v of DMSO.

10. The method according to claim 8 , wherein the vacuum tube is filled with about 10% v/v of DMSO.

11. The method according to claim 8 , further comprising incubating the cells mixed with DMSO in the vacuum tube before freezing.

12. The method of claim 8 , wherein the connector is a Luer, a fluid fitting used for making leak-free connections between a male-taper fitting and its mating female part.

13. A method for cryopreservation of cells from a bioreactor, the method comprising,
cultivating cells in the bioreactor as a perfusion culture to 10-200 MVC/mL,
providing an access device having a needleless connector configured for forming a needleless connection to the bioreactor;
using the needleless connector of the access device to form a needleless connection with the bioreactor to access the cultivated cells;
sampling or harvesting the cells from the bioreactor by inserting a vacuum tube provided with 5-20% v/v of freeze media into the access device such that the cells are instantly mixed with the freeze media; and
freezing said vacuum tube for cryopreservation of the cells.

14. The method according to claim 13 , further comprising adding a freeze media into an empty vacuum tube to form the vacuum tube provided with freeze media, wherein the freeze media is DMSO, glycerol or methyl cellulose.

15. The method according to claim 13 , wherein one or more steps of the method are automated.

16. The method according to claim 13 , further comprising incubating the cells mixed with freeze media in the vacuum tube before freezing.

17. The method according to claim 13 , wherein the vacuum tube is filled with about 10% v/v of DMSO.

18. The method according to claim 13 , wherein the access device comprises a needle for inserting into the vacuum tube.

Assignees

• Cytiva Sweden Ab

Inventors

• Faldt Eric

Classifications

• B01L3/50825 Primary

  Closing or opening means, corks, bungs · CPC title

• A01N1/147

  Carriers for immersion in cryogenic fluid for slow freezing or vitrification · CPC title

• A01N1/122 Primary

  Preservation or perfusion media · CPC title

• F25D2303/0842

  inside the beverage contained in a bottle, can, drinking glass, pitcher or dispenser · CPC title

• F25D3/08

  portable, i.e. adapted to be carried personally · CPC title