Substrate unit, preservation device and method for the cryopreservation of a biological sample

Patent metadata

Field	Value
Publication number	US-9781918-B2
Application number	US-201314412615-A
Country	US
Kind code	B2
Filing date	Jul 1, 2013
Priority date	Jul 4, 2012
Publication date	Oct 10, 2017
Grant date	Oct 10, 2017

Abstract

A substrate unit for cryopreservation of a biological sample includes: a substrate platform having on a front side thereof a cultivation surface for receiving the biological sample; a first chamber including the cultivation surface of the substrate platform, wherein the first chamber is configured for receiving a cultivation liquid; a second chamber configured for receiving a temperature control medium; and a chamber bracket for receiving the substrate unit in a cryopreservation device in a pivotable manner, wherein the first and second chambers are connected to each other in an adjacent manner, and the substrate platform forms a separating wall between the first chamber and the second chamber, and wherein the back side of the substrate platform faces the second chamber. The invention further relates to a cryopreservation device, wherein the substrate unit is arranged in a rotatable manner, and a method for the cryopreservation of a biological sample.

First claim

The invention claimed is:

1. A substrate device for the cryopreservation of a biological sample including biological cells, comprising
a substrate platform having a front side and a back side, the front side of the substrate platform forming a cultivation surface for receiving the biological sample, and
a first chamber, in which the cultivation surface of the substrate platform is included, the first chamber being configured for receiving a cultivation liquid, wherein the first chamber is separated from surroundings by a cover,
a second chamber, which is configured for receiving a temperature control medium, and
a chamber holder, which is configured for receiving the substrate device in a cryopreservation apparatus in a pivotable manner, wherein the chamber holder comprises supporting elements being arranged in a plane parallel to an extent of the substrate platform and defining a lateral axis of rotation, which supporting elements can be pivotably coupled with an external support, wherein
the first chamber and the second chamber are connected to each other in an adjacent manner,
the substrate platform forms a separating wall between the first chamber and the second chamber, the back side of the substrate platform facing towards the second chamber and forming a liquid-tight closure with the second chamber, and
the first chamber and the second chamber are configured such that the biological sample in the first chamber is isolated from the temperature control medium in the second chamber, so that there is no exchange of substances between the first chamber and the second chamber.

2. The substrate device according to claim 1 , wherein
the back side of the substrate platform is exposed in the second chamber, and
the substrate platform is formed with a thickness and a thermal conductivity which, when the back side of the substrate platform is wetted with a first temperature control medium at a temperature below −120° C., allow vitrification of the biological sample on the cultivation surface.

3. The substrate device according to claim 1 , wherein the substrate platform has at least one of the following features:
the substrate platform has a thickness below 200 μm,
the substrate platform is made of glass, plastic, semiconductor material or metal, and
the substrate platform is made of a transparent material.

4. The substrate device according to claim 1 , wherein
the substrate platform is detachably connected to the first or second chamber via a liquid-tight substrate holder.

5. The substrate device according to claim 1 , which has at least one of the following features:
the substrate platform is connected to the first or second chamber via a compensating section, the compensating section being configured for absorbing temperature-dependent stresses between the substrate platform and the first or second chamber, and
the first chamber has a pressure equalizing valve which is configured for equalizing the pressure between the first chamber and its surroundings.

6. The substrate device according to claim 1 , wherein
the substrate platform is an integral component of the first or second chamber.

7. The substrate device according to claim 1 , which comprises
a delivery device, which is configured for delivering at least one of the cultivation liquid and/or the temperature control medium.

8. The substrate device according to claim 7 wherein
the delivery device comprises at least one of at least one media line and a microfluidic unit.

9. The substrate device according to claim 1 , wherein
the first and second chambers are detachably connected to each other.

10. The substrate device according to claim 9 wherein
the second chamber is firmly connected to a chamber frame, which is configured for detachably receiving the first chamber.

11. The substrate device according to claim 9 , wherein
the first chamber is connected to the second chamber via a screw joint.

12. The substrate device according to claim 1 , wherein
the first chamber comprises several sub-chambers which are arranged adjacent to the second chamber, the substrate platform forming a separating wall between the sub-chambers and the second chamber.

13. The substrate device according to claim 2 , wherein
the backside of the substrate platform is wetted with liquid nitrogen.

14. A cryopreservation apparatus, comprising
at least one substrate device according to claim 1 , and
a rotating device, which is configured for receiving the at least one substrate device, wherein
the substrate device, by the rotating device being capable of pivoting between a cultivation state, in which the substrate platform forms the floor of the first chamber, and a temperature control state, in which the substrate platform forms the floor of the second chamber.

15. A method for the cryopreservation of a biological sample containing biological cells, using a substrate device according to claim 1 , comprising the following steps:
providing the biological sample on the cultivation surface of the substrate platform in the first chamber, the biological cells being surrounded by a cultivation liquid, and
receiving a cooling medium into the second chamber, which is adjacent to the first chamber and is connected to the latter in such a way that the substrate platform forms the separating wall between the first chamber and the second chamber, the temperature of the substrate platform being lowered and the biological sample being converted to a frozen state, wherein

the cultivation of the biological cells comprises the following steps:
providing the first chamber and the second chamber in a cultivation state, in which the substrate platform forms the floor of the first chamber and the cultivation liquid is filled into the first chamber, and
receiving the biological cells in the adherent state on the cultivation surface of the substrate platform, wherein
the first chamber and the second chamber are pivoted into a temperature control state, in which the substrate platform forms the floor of the second chamber, and the cultivation liquid flows out of the first chamber, before the cooling medium is received in the second chamber.

16. The method according to claim 15 wherein the cultivation of the biological cells comprises the following steps:
providing the first chamber and the second chamber in a temperature control state, in which the substrate platform forms the floor of the second chamber, and
receiving the biological cells in droplets of the cultivation liquid, which are suspended from the cultivation surface of the substrate platform.

17. The method according to claim 15 , wherein
the temperature of the substrate platform is lowered at a cooling rate such that the biological sample is vitrified.

18. The method according to claim 15 , wherein
the biological sample includes biological cells of different types, which are subjected to a common cultivation on the cultivation surface.

19. The method according to claim 15 , comprising the following step:
receiving a heating medium into the second chamber, the temperature of the substrate platform being raised and the biological sample being converted to a thawed state.

Assignees

• Fraunhofer-Gesellschaft Zur Foerderung Der Angewandten Forschung E V

Inventors

• Zimmermann Heiko
• Beier Axel
• Neubauer Julia
• Fuhr Guenter R

Classifications

• C12M45/22 Primary

  Means for packing or storing viable microorganisms (casings for storing cell samples A61B10/0096, preservation of living parts of the human or animal body A01N1/10) · CPC title

• A01N1/021 Primary

  Human Necessities · mapped topic

• A01N1/122 Primary

  Preservation or perfusion media · CPC title