DK201500102U4

DK201500102U4 - Apparatus for cell detachment

Info

Publication number: DK201500102U4
Application number: DKBA201500102U
Authority: DK
Inventors: Ivan Svendsen; Thomas Andreasen
Original assignee: Thomas Andreasen; Ivan Svendsen
Priority date: 2015-07-16
Filing date: 2015-07-16
Publication date: 2016-10-28

Abstract

The generation solves the problem of releasing cells from the surface of cell culture vessels by applying ultrasonic energy at right angles to the outside of the cell culture surface. The container is held in a fixture that holds the container tightly between a pretensioning and a sonotrode. This ensures that the cell culture vessel is maintained in a certain position relative to the sonotrode, at the same time ensuring that the sonotrode and the cell culture vessel is brought into close contact with each other with a defined force. The vibrations generated by the ultrasonic energy release, releases the cells from the surface. This method is faster and more gentle to the cells over the prior art technologies.

Description

Title

An apparatus for releasing cells

Frembringelsens scope

The generation provides a simple and effective way to release the cells from the surface of rigid cell culture containers, such as T-flasks.

Background art

Traditional releasing the adherent cells from a cell culture surface in connection with the passage of the cells is carried out by treating the cells with enzymes as trypsin, chymotrypsin, collagenase, or other proteases. Enzymes can, however, damage to the cells or cell surface proteins, and result in a risk of contamination of cells with viruses for example.

Another method of release of the cells is by mechanically scraping the cells of the cell culture surface with a suitable scraper. This method can be very harmful for the cells and result in low viability. The cell scraping and the release of enzymes are both multi-step techniques that are time consuming. US Patent 8,114,646 describes a method for release of cells from the multi-layer cell culture containers by application of ultrasonic energy, but the system is unable to release the cells from rigid surfaces, as described in column 10 lines 54-57. W02014134090 describes a different principle, where ultrasonic energy is used. This patent is based on "relative scan motion" to free up sufficient cells from around the cell growth area. This is a very complicated way to try to release cells. Furthermore, pre-treatment with EDTA required to obtain a satisfactory result.

The technical problem to be solved

This generation solves the problem of releasing cells from the cell culture containers having a rigid cell culture surface, in a simple and effective manner, without the use of enzymes, EDTA or cell scraper methods and without cells, or cell surface proteins damaged.

The new technique

The generation of an apparatus for releasing the cells which are new in that a cell culture vessel (6) into a removable fixture (5) and is clamped between a sonotrode (4), which is connected to an ultrasonic generator (9), which is placed in a cabinet (1), and a pretensioning unit (3), which is mounted in a hinged lid (2) with a lock (7), wherein the sonotrode is brought to sit at right angles to the cell culture surface at a fixed position. The apparatus include a timer (8), which can sit anywhere on the device.

The problem to dislodge the cells from the surface of the cell culture container, has been solved by feeding cell culture container's outer surface ultrasound energy perpendicular to the cell culture surface. The cell culture container is held in place in a fixture and is clamped between a pretensioning unit and a sonotrode. This ensures that the cell culture vessel is maintained in a certain position relative to the sonotrode, at the same time ensuring that the sonotrode and the cell culture vessel is brought into close contact with each other with a defined force. The vibrations arising from the ultrasonic energy release, loosen the cells from the surface. This method is faster than prior art techniques.

The technical effect

By using ultrasonic energy to release the cells from the culture surface, presents this generation a quick and effective way to solve the problem. Furthermore, this generation requires no prior treatment of the cells with EDTA or enzymes. The method is more rapid than traditional methods and does not damage either cells or cell surface proteins. The result is higher cell viability compared to cell scraping or enzymatic treatment.

List of Figures

Figure 1 shows the subject matter, with the lid partially open

Figure 2 shows a vertical section of the subject matter, with the lid partially open

Figure 3 shows a vertical section of the subject matter, with the lid closed;

Titles 1 - Housing 2 - Hinged lid 3 - pretensioning 4 - sonotrode 5 - Fixture 6 - Cell Culture Container 7 - Handle with lock 8 -Timer 9 - Ultrasonic Generator 10 - Pressing shoes

Experimental Example

A particularly advantageous embodiment of the subject matter shown in Figures 1 to 3.

The execution consists of a cabinet (1) with a hinged lid (2). The lid contains a pretensioning unit (3) having a pressing shoe (10) and a handle with a lock (7). The cabinet contains an ultrasound generator (9) with a sonotrode (4) and a timer (8) which determines the duration of ultrasonic exposure. The cell culture flask (6) is placed in a fixture (5). The fixture could be replaced to suit different types of cell culture bottles. The fixture ensures that the bottle remains in a particular position relative to the sonotrode. The prestressing of the cover ensures a close contact between a number of different cell culture flasks, and the sonotrode. In this design, the production, the user can put cell culture bottle in the fixture, close and lock the lid, whereby the prestressing press bottle tightly against sonotrode. The user can then make the duration of ultralydspåvirkingen, based on the cylinder and cell type, typically from 5 to 30 seconds, but the duration may be longer if necessary. The cells are then released and is ready to be passed to the new cell culture bottles.

Claims

1.

An apparatus for detaching of cells from rigid cell culture surfaces which are new by a cell culture vessel (6) into a removable fixture (5) and is clamped between a sonotrode (4), which is connected to an ultrasonic generator (9) by is located in a cabinet (1), as well as a pretensioning unit (3), which is placed in a hinged lid (2) with a lock (7), wherein the sonotrode is brought to sit at right angles to the cell culture surface at a fixed position.

The apparatus include a timer (8), which can sit anywhere on the device.

2. An apparatus for detaching of cells according to claim 1, which is new one pre-tension device (3) has a pressure shoe (10) mounted on a rod at its end that affects cell culture vessel (6). 3. An apparatus for detaching of cells according to one of the preceding claims, which is new in that the lid (2) hinged along one side to be added to that by the lever rule, from a force which pre-tension device (3) operates by. 4. An apparatus for detaching of cells according to one of the preceding claims, which is new, by the sonotrode (4) may have more than one point of contact with the cell culture vessel (6).