PRODUCT AND METHOD FOR CELL PRESERVATION

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.

Claims

1. A product for cryopreservation of cells, comprising a vacuum tube provided with freeze media, and a penetratable cap on said tube.

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

3. The product according to claim 1, wherein the vacuum tube is filled with 5-20% v/v, preferably 10% v/v freeze media.

4. The product according to claim 1, comprising an access device for receiving said vacuum tube and provided with a needle to be inserted into the vacuum tube and provided with a connector for connection to a cell source, such as a cell culture in a bioreactor.

5. A method for cryopreservation of cells, comprising sampling or harvesting cells into a vacuum tube provided with freeze media; closing said tube with a penetratable cap; and freezing said tube.

6. The method according to claim 5 comprising cultivating cells in a bioreactor, harvesting cells from said bioreactor into a vacuum tube provided with freeze media, wherein the cells are instantly mixed with the freeze media; removing said tube from said bioreactor, and freezing said tube and its content.

7. The method according to claim 5, wherein the freeze media is DMSO, glycerol or methyl cellulose.

8. The method according to claim 5, wherein the vacuum tube is filled with 5-20% v/v, preferably 10% v/v freeze media.

9. The method according to claim 5, wherein the cells are cultivated as a perfusion culture.

10. The method according to claim 5, wherein the cells are cultivated to 10-200 MVC/mL.

11. The method according to claim 5, wherein one or more steps are automated.

12. The method according to claim 5, wherein the cells are incubated before freezing.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0017] FIG. 1A shows a schematic view of a vacuum tube prepared with concentrated cryopreservation liquid.

[0018] FIG. 1B shows a schematic view of an access device for receiving the vacuum tube and having a needle for penetrating the lid of the vacuum tube.

[0019] FIG. 1C shows a schematic view of the access device connected to the vacuum tube and ready to be inserted to the bioreactor for cell sampling.

DETAILED DESCRIPTION OF THE INVENTION

[0020] The present invention will now be described in relation to a non-limiting example and the accompanying figure.

[0021] FIG. 1A shows a schematic view of a vacuum tube (1) prepared with concentrated cryopreservation liquid (2). The tube is also provided with a penetratable and removable cap (3) and penetratable septum (4) of silicon or other soft material. The tube may be any vessel appropriate for cryo preservation applied with vacuum, for example tubes may be of glass or plastic.

[0022] FIG. 1B shows a schematic view of an access device (5) comprising a protective housing (5) for receiving the vacuum tube and having a needle (6) for penetrating the lid of the vacuum tube. The needle is provided with a protective sleeve (7) of rubber or silicon. The access device is provided with a Luer or other connection to a bioreactor for cell sampling directly into the vacuum tube.

[0023] FIG. 1C shows a schematic view of the access device (5) connected to the vacuum tube (1) and ready to be inserted to the bioreactor via the connection (8) for cell sampling. The cryo vessel is pushed into the protective cap (3) along the direction of the arrows. The needle will penetrate the soft material septum. The vacuum will withdraw the cell broth from the bioreactor for mixing directly with the cryopreservation liquid (freeze media). Thereafter, the tube is placed in a fridge for cell banking.

EXAMPLE

[0024] Conventional 5 ml vacuum tubes are filled with 0.5 ml concentrated DMSO (freeze media) using a syringe. A CHO (Chinese Hamster Ovary) culture is brought to up to 10-100 MVC/mL, preferably using perfusion culture. The access device with sleeve protected needle (6) is attached to the bioreactor via the connector (8). Cryo-liquid prepared vacuum tubes according to the invention are used to draw cell samples from the bioreactor. The vacuum tubes are inserted in the access device and the vacuum draws the cells broth into the tube through the needle of the access device. The cell broth and the freeze media are instantly mixed.

[0025] Thereafter the filled cryo prepared vacuum tubes are directly transferred to a cell banking fridge. If necessary, the cells in the tubes are shortly incubated before freezing.

[0026] The procedure to draw cell samples from bioreactors does not damage the cells and is very fast which enables production of large cell banks with minimum of open handling. If desired, the process may be automated.