CZ2009264A3 - Method of addition, removal or replacement of liquid medium when culturing biological cells and culturing device - Google Patents

Abstract

A method of adding, removing or replacing a liquid medium in culturing biological cells without any contact with these cells, and a culture device (1) for culturing biological cells and manipulating said cells, preferably embryonic cells, containing at least one elongated culture well (6) with a cross section V-shaped and at least one well (7) for medium exchange, which allows culturing embryos in groups in close proximity using the beneficial effects of the growth factors released by the embryos, ensuring full identification of each individual embryo throughout the cultivation period and allowing non-contact exchange of nutrient medium. The device (1) can be arranged on a common base (11) in combination with another device (12), such as for in vitro fertilization, and covered with a lid (10).

Description

A method of adding, removing, or replacing a liquid medium in a culture of biological cells and a culture device

Technical field

The invention relates to a method of adding, removing or replacing a liquid medium in the cultivation of biological cells and a cultivation device for cultivating and manipulating biological cells, preferably embryonic cells.

BACKGROUND OF THE INVENTION

Currently, human embryos and other animal embryos are cultured in vitro in various culture equipment or multi-well plates.

Such a typical arrangement is a conventional flat device, at the bottom of which an aqueous culture medium in the form of individual drops is placed. The drops are usually covered with mineral oil during cultivation. Extreme care should be taken to prevent droplet displacement when handling equipment.

US 5,891,712 discloses an improved microdrop culture device in which a flat bottom is embedded with a plurality of circular wells with open top surfaces, where the wells may have marks to identify individual wells for monitoring purposes. According to the disclosure, the culture device together with the removable lid is capable of providing a stable environment for gametes / embryos development and thus a shorter incubation time for the procedures, clearly identifying the samples contained therein, and preventing displacement of the droplets during handling of the devices.

The embryos can be grown for up to five to six days, during which time the culture medium needs to be changed to prevent the accumulation of toxic products and the depletion of nutrients.

In any of the systems used, oocytes / embryos are transferred to another compartment (chamber, well, etc.) or simply to another drop of fresh medium. During this manipulation, clusters of embryos are separated, reducing the supportive effect of autocrine / paracrine growth factors that release the embryos. In addition, they must be handled by qualified personnel using glass / plastic devices and thus embryos are exposed to damage and / or loss. In order to compensate for the loss of embryos during handling, the initial number of embryos must always be greater than the number ultimately implanted in the woman. However, such excess embryos should be avoided for ethical reasons.

Therefore, it would be highly desirable to have an improved culture device 10 as well as a corresponding improved method suitable for noncontact nutrient media exchange, wherein cultured embryos / cells are neither tampered with nor come into contact with media exchange devices (eg, pipettes).

Several embryo culture techniques are routinely used. In any of the 15 methods used, the viability of an individual embryo is assessed by light microscopy based on morphological features. This requires continuous identification of each embryo from the group throughout the culture period. Cultivation in clusters is inappropriate because embryos can easily be damaged during handling. To avoid this disadvantage, the individual embryos are placed in culture dishes at spaced apart locations. However, cultivation of individual embryos reduces the influence of growth factors released by the embryos that support their development.

In addition, during culture up to five days, nutrient medium replacement is required, in which case embryos must be transferred to fresh medium and the by-products of growth are lost. If embryos are not transferred, the accumulation of toxic metabolites in the culture medium may impair further embryo development.

US 6,448,069 discloses an embryo culture device comprising at least 30 one partitioned structure, wherein each individual compartment is of a suitable size for receiving one embryo, and wherein fluid migration between compartments is allowed. In a preferred embodiment, each compartment is defined by spaced apart spikes pointing upwardly from the bottom of the well or by a liquid permeable membrane. After placing the embryos in compartments in each structure, each structure is covered with a drop of nutrient medium, so that embryos in each divided structure can share by-products with other embryos in the same structure. However, there is still a need for contact handling of the embryos when transferred to fresh media.

Thus, there is still a need for a simple and inexpensive cultivation device which, without creating more complicated microstructures such as spaced spikes or a liquid permeable membrane, can cultivate embryos in groups in close proximity to each other using the supportive effects of embryo-released growth factors in which ensure full identification of each individual embryo throughout the cultivation period, while allowing non-contact exchange of the nutrient medium.

SUMMARY OF THE INVENTION

These problems are solved by the first object of the present invention, which is a method of adding, removing or replacing a liquid medium in cultivation and / or any other manipulation of biological cells, preferably embryonic cells housed in one or more wells arranged in the bottom wall of the culture device. manipulating the cells or contacting the media handling device, such as a pipette, with said cells.

This method allows any handling of any liquid aqueous or non-aqueous media, such as nutrient media, as well as wash, storage, treatment, buffering, equilibration, protective or covering media that should be brought into contact with the cells.

In this method of the invention, any manipulation, such as transferring cells or embryos to fresh medium by a pipette, is avoided, and thus disruption of embryo clusters can be avoided.

The method of the invention comprises two basic steps:

a step of adding medium to the culture device, wherein in this step the level of the medium in the culture device gradually increases above the open ends of said wells, thereby establishing a fluid communication between the added medium and the interior of said wells 5 through the open ends of said wells; or a step of removing the medium from the culture device, wherein in this step the level of the medium in the culture device gradually decreases below the open ends of said wells, thereby breaking the connection through 10 liquid open ends of said wells between the added medium and the inner space of the wells.

Although the method of the invention allows for the addition or removal of media from any culture device having biological cell culture wells (culture wells), for example, from any 15 location on the bottom wall of the culture device in which the media handling device does not contact the cells preferably from the lowest point of the upper surface of the lower wall of the cultivation device, more preferably the wells arranged at the lowest point of the upper surface of the lower wall of the cultivation device which contains no cells preferably, the medium is added to or removed from the medium change well of a special culture device, which is a further object of the present invention.

In this most preferred embodiment, the medium exchange method begins by aspirating the medium, if present, from the medium exchange well disposed at the lowest point of the upper surface of the bottom wall of the culture device.

Surprisingly, the inventors have found that after removal of the medium, the one or more wells in which the cells / embryos are stored (culture wells) remain not only filled with the medium, but the embryos are not disturbed or raised and maintain their original order.

When adding fresh medium, preferably by extruding the medium into the medium exchange well arranged at the lowest point of the upper surface of the bottom wall of the cultivation device, the medium level gradually rises above the open ends of said cultured embryo / cell culture wells and the original medium is extensively diluted with fresh media.

Furthermore, the operation can be performed very easily and simultaneously in one step for all wells with cultured embryos / cells, is less time consuming and has less demands on experienced laboratory staff than prior art methods. In addition, no fine embryo / cell manipulation tools are required.

The dilution ratio and hence the possibility of maintaining the embryo / cell released growth factors in their close proximity may be affected to some extent by the amount of fresh medium added and thus the level of fresh medium above the open ends of said cultured embryo / cell culture wells and incubation time before medium.

In the step of removing the medium from the culture device, the level of the medium in said culture device gradually decreases below the open ends of said culture wells, thereby interrupting the liquid-open connections of the culture wells between the added medium and the inner space of said wells. as standard cultivation in microdroplets.

If desired or necessary, by omitting the above media removal step from the culture device, the level of the media in said culture device remains above the level of the open ends of the wells, and fluid communication between all wells of the culture device through the medium is maintained.

If desired or necessary, the above media removal and addition steps may be repeated several times, for example, to achieve embryo / cell washing.

In addition, the method of the invention allows for the addition or removal of a medium under a layer of a non-aqueous liquid coating such as mineral oil as commonly used in the art.

In a preferred embodiment, the medium is removed by aspiration from a well arranged at the lowest point of the upper surface of the bottom wall of the culture apparatus, under a layer of non-aqueous liquid so that substantially all of the non-aqueous liquid remains in the apparatus. The addition of fresh medium in this case is preferably done by forcing the medium into the same well, also under a layer of non-aqueous liquid covering. In this way, the non-aqueous liquid may remain unchanged throughout the cultivation.

Preferably, the method is carried out on any embodiment of the culture device as described below.

Another object of the present invention is to provide a cultivation device comprising a bottom wall and a side wall defining an interior space of the cultivation device, wherein at least one elongated cultivation well is embedded in the bottom wall, viewed along its long axis with a V-shaped cross section. and at least one medium exchange well directed with its upper open end into the interior of the device.

It has been found that embryos cultivated in a series of elongated culture wells having a substantially V-shaped cross section and thus forming a groove or channel-like structure in the bottom wall of the culture device retain their original position throughout the entire culture process.

The culture wells are designed with appropriate depths and walls with a suitable angle to minimize embryo flotation during handling while maintaining good accessibility to the embryos during handling.

For the non-contact addition, removal or replacement of the medium, at least one medium exchange well 30 is recessed in the bottom wall of the culture device, with its upper open end directed into the interior of the culture device.

Preferably, at least a portion of the upper surface of the bottom wall of the culture device comprising one or more culture wells has a gradient towards the media exchange well for easy collection of media in the media exchange well.

The bottom of the culture well is preferably horizontal. In two or more culture wells, the bottoms are preferably at the same horizontal level.

The edge of the upper open end of the culture well facing the interior of the culture device may be raised above the upper surface of the bottom wall of the culture device to form an edge with a horizontal upper end, said edge surrounding the upper open end of the culture well. This edge ensures a constant depth of the elongated culture well along its longer axis and, in addition, minimizes embryo buoyancy during medium change and allows the worker to maintain equal access to all embryos during handling. In the case of two or more culture wells, the horizontal upper ends of these edges are preferably at the same horizontal level.

Thus, one media exchange well may form a "unit with one or more surrounding culture wells, wherein a portion of the upper surface of the bottom wall containing such a" unit "may have a slope toward that media exchange well. A single culture device may comprise several such separate units at any point of the bottom wall, each unit being separated from the other units by side walls, or wherein two or more units may be surrounded by a single common side wall. Such an arrangement is useful, for example, in genetic analysis to cultivate individual embryos.

The longer axes of the culture wells surrounding a single media exchange well are preferably parallel to each other to allow easy access to all embryos by a pipette held at the same angle with respect to the culture device of all culture wells, as well as safe identification of the embryos.

The shape of the media change well is not critical. In a preferred embodiment, the media exchange well has a tapered side wall with a wider upward end, to allow removal of even the smallest amount of media remaining, and to easily distinguish the level of non-aqueous coating liquid when used. The media change well may have a round or flat bottom.

In order to ensure the same medium exchange conditions in the individual culture wells, the culture wells are preferably arranged around the central media exchange well at substantially the same distance from the central media exchange well.

The bottom wall of the culture device preferably comprises markings, such as numbers or letters, to identify individual culture wells.

The corner at the point of contact between the upper surface of the bottom wall and the side wall is preferably rounded or tapered into the interior of the cultivation device to minimize the amount of liquid adhering.

The culture device may further comprise a removable lid covering the interior space of said culture device.

It is possible for the culture device to be arranged on a common base with one or more identical culture devices and / or one or more other devices or structures, such as reservoirs for replacement medium or other auxiliary structures, which may be separated from each other by the side walls.

Such an additional structure may be an in vitro fertilization oocyte section known in the art, including separate sperm cells / chambers or eggs, connected at their upper ends through a narrow channel. After leaching, fertilized eggs can only contact live and highly mobile sperm.

Any such structure may include a removable lid.

In the case of arranging multiple devices and / or structures of the same or different kind on a common base, it may be advantageous if the individual lids overlapping the interior of each cultivation device and / or the interior of any other device present on the common base are replaced by a single common lid overlapping all the devices or structures on the same common base.

Advantageously, the common lid, in its first position, overlaps all the cultivation devices and / or other devices arranged on the common base, and in its second, horizontally rotated position relative to the first position, allows access to the interior space of said devices arranged on the common base, e.g. segments or slices. This principle has already been applied in the art, for example in a well plate with a cover plate according to FI884551.

BRIEF DESCRIPTION OF THE DRAWINGS

Giant. 1 illustrates a side section of one embodiment of a culture device 1 according to the invention with one central well 7 for media exchange and two culture wells 6;

Fig. 2 illustrates a top view of the cultivation apparatus 1 of Fig. 1 with one central well 7 for media exchange and four cultivation wells 6 arranged around the central well 7 for media exchange;

Fig. 3 illustrates a top view of a cultivation device 1 with one central well 7 for changing media and four cultivation wells 6 arranged around the central medium changing well 7 containing the markings A, B, C and D on the bottom wall 2 identifying the culture wells 6;

Fig. 4 illustrates a side cross-section of the embodiment of Fig. 1 in which the upper surface 8 of the bottom wall 2 of the cultivation device 1 has a slope towards the medium change well 7, with an optional lid 4 overlapping the inner space 5;

FIGS. 5A-C illustrate a side sectional view of several embodiments of the culture well 6 viewed along the longer axis of the well 6, optionally showing the embryo 9 present in the well 6;

Fig. 5D illustrates a side sectional view of the culture well 6 viewed along the long axis of the well 6, showing an edge 13 surrounding the upper open end of the culture well 6;

Fig. 5E illustrates a side section of the culture well 6 viewed perpendicularly to the longer axis of the well 6, showing an edge 13 surrounding the upper open end of the culture well 6;

Fig. 5F illustrates a detail view from above of the culture well 6;

Fig. 5G illustrates the same view as Fig. 5F and shows embryos 9 placed in culture well 6;

FIG. 6 illustrates a side section through one embodiment of a media change well 7;

Fig. 7 illustrates a top view of an arrangement of two culture devices 1, each comprising four wells 6 around one central well 7 for changing media, on a common base 11;

Figures 8A-B illustrate two different oblique views of a cultivation device 1 arranged on a common base 11 in combination with one in vitro fertilization device 12, with one possible embodiment of a single removable lid 10;

Figures 9A-B illustrate two different oblique views of a cultivation device 1 arranged on a common base 11 in combination with one in vitro fertilization device 12, with the lid 10 in the open position (A) and rotated to its closed position (B).

DETAILED DESCRIPTION OF THE INVENTION

Certain preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings.

It should be understood by those skilled in the art that although embodiments of the present invention are described in the specification on embryos, the invention is not limited to embryonic cells of humans or other animals, but may be applicable to all sensitive and / or fragile biological materials such as oocytes and oocytes. other animal or plant cells or parts of cells such as protoplasts.

In addition, the terms "culture well" or "cultivation" should be interpreted to include all conceivable means such as receiving, maintaining, preserving, washing, inoculating or processing biological materials.

The non-contact media change according to the first embodiment of the present invention can be used essentially to add or remove media from any culture device having biological cell culture wells (culture wells). However, the process is preferably carried out in a culture device according to the invention.

In the contactless exchange of the medium of the invention, the cultured embryos / cells are neither manipulated nor come into contact with media exchange devices (e.g. pipettes), i.e. the nutrient medium is preferably exchanged by removing it from / adding it to the media exchange well 7. In a typical cultivation procedure, the following steps are performed:

Step 1 - introduction of nutrient medium

In this step, the media change well 7 is filled using, for example, a conventional replaceable tip pipette with nutrient medium. The level of the medium in the culture device 1 is thus gradually raised above the level of the open upper ends of the culture wells 6 and finally the medium is overlaid with mineral oil.

Step 2 - Add embryo

In this step, embryos 9 are pipetted into one or more V-shaped culture wells 6. This handling may require special handling tools and trained personnel.

It is possible to cultivate the embryos / cells in this arrangement, i.e. with the level of the medium left above the level of the open upper ends of the culture wells 6, or proceed as needed with step 3.

Step 3 - Remove excess media

In this step, the suction device (pipette tip) is inserted into the medium change well 7 and the medium is aspirated. After this operation, only the culture wells 6 containing the embryos remain filled with the medium and the medium remains covered with mineral oil to prevent evaporation of the medium and changes in its osmoiarity.

Step 4 - Replace the spent media

In this step, the aspirator (pipette tip) is inserted into the media change well 7 and the new medium is expelled from the pipette until the medium level in said culture device 1 rises above the level of the upper ends of the wells 6, thereby establishing a fluid communication between with the added medium and the interior of the wells covered with fresh medium over the open ends of said wells 6 containing embryos 9. The spent medium in the culture wells 6 is extensively diluted with fresh medium.

The medium may remain covered with the same mineral oil throughout the cultivation period to prevent evaporation of the medium and changes in its osmotic pressure.

One embodiment of a simple culture device 1, according to a second embodiment of the present invention, having a bottom wall 2 and a side wall 3 defining an interior space 5 of the culture device 1, comprises one media exchange well 7 and two culture wells 6 embedded in the bottom wall 2 of said device 1 The apparatus shown in Fig. 1 is formed of a sufficiently strong and transparent material such as polystyrene.

A perspective view of a similar culture apparatus 1 is shown in Figure 2. In this arrangement, it is represented by one central media exchange well 7 surrounded substantially equidistantly by four culture wells 6, with the longer axes of all four culture wells 6 running parallel to each other.

In order to allow easy access to cells by manipulation devices such as a pipette, the diameter of the interior 5 of the culture device 1 suitable for in vitro embryo culture (or the diameter of portions of the upper surface 8 of the bottom wall 2 surrounding the media exchange wells 7 containing one or more culture wells 6) can be, for example, 15 to 30 mm. The depth of the interior space 5 of the cultivation device 1 can be, for example, 5 to 15 mm, without being limited to these dimensions.

A top view of a similar culture apparatus 1 with numbers identifying each of the culture wells 6 is shown in Figure 3.

An embodiment showing that the upper surface 8 of the bottom wall 2 of the cultivation device 1 has a gradient to the media exchange well 7 in the area surrounding the media exchange well 7 is shown in Fig. 4. The angle β of the media exchange well 7 is preferably 0.5 to 2 °, which is sufficient for gravitationally draining the medium into the medium change well 7, but may range from 0 ° (flat top surface 8) to, for example, 10 °.

It should be noted that the medium change well 7 does not necessarily have to be located in the central region of the culture device. This embodiment is shown in Fig. 7, in which the combined culture apparatus comprises two media change wells 7 on a common base 11, and thus two separate portions of the upper surface 8 of the bottom wall 2 surrounding said media change wells 7 and comprising one or more culture media. the wells 6 have a slope to the two respective medium exchange wells 7. Moreover, if the two parts of the surface 8 of the bottom wall 2 are surrounded by the side walls 3, two separate cultivation devices L are formed on the common base 11.

The elongated culture well 6 has a substantially V-shaped cross section as viewed along its longer axis, as shown in Fig. 5, thereby forming a groove or channel structure in the bottom wall 2 of the culture device 1 in which the embryos 9 retain their original positions during of the entire cultivation process.

The term "substantially V-shaped cross-section" is intended to mean not only the embodiment shown in Fig. 5A but also the flat bottom cross-section shown in Fig. 5B, or any other cross-section with a wider upward end as long as it keeps the embryos protected a stable, permanent order to identify embryos throughout the culture period.

For this purpose, the possible flat part of the bottom "V" should not be wider than the diameter of the embryo or any other cultured cell, for example 0.02 to 0.08 mm. The volume of the culture wells 6 depends on the cultured embryos / cells and may, for example, be approximately 10 to 50 μΙ.

A suitable angle α between the longitudinal walls of the elongated culture wells to minimize embryo buoyancy during handling, while maintaining good accessibility during handling of cultured embryos from all directions, may be 40 to 60 °, and the bottom length of the well 6 preferably calculated on culture For example, 10 to 15 embryos in a row are equal to 10 to 20 times the embryo diameter (typically 0.015 mm), i.e. preferably about 3 mm.

Embryos placed in elongated culture well 6 are shown in Figure 5G.

The suitable angle of the transverse walls shown in Fig. 5E of the elongated culture wells 6 is not critical and preferably is the same as the angle α of the longitudinal walls of the elongated culture wells 6.

As also seen in Fig. 5E, the bottom of the culture well 6 is preferably horizontal. In two or more culture wells, the bottoms are preferably at the same horizontal level - see Figures 1 and 4.

The edge of the upper open end of the culture well 6 facing the interior 5 of the culture device 1 is preferably raised above the upper surface 8 of the bottom wall 2 of the culture device to form an edge 13 as shown in FIG. the edge 13 surrounds the upper open end of the culture well 6. The width of the edge as well as its height above the top surface 8 may preferably be, for example, 0.2 to 2 mm.

It will be appreciated that in the case of the upper surface 8 of the bottom wall 2 with the slope towards the central media exchange well 7, the height of the rim 13 above the upper 30 surface 8 is not the same.

In the case of two or more culture wells 6, the horizontal top ends of said edges 13 are preferably at the same horizontal level.

In the preferred embodiment shown in FIGS. 1 to 4, the medium change well 7 has a larger volume and a greater depth compared to the embryo culture wells 6. The volume of the media change well 7 may be approximately 50 to 150 μΙ. The media change well 7 preferably has a conical shape with a wider upward end, and may, for example, have a round or flat bottom to be readily accessible by pipette and / or liquid handling device in the event of automatic media change.

The angle γ of the side walls of the medium change well 7 is shown in Fig. 6 and is selected such that the level of the medium or the interface between the medium and the covering non-aqueous liquid is clearly visible on the side walls and preferably equal to the angle γ of the longitudinal walls of the elongated culture wells. 6.

The device 1 may be covered by a removable lid 4 as shown in Fig. 4, allowing gas exchange between the interior space 5 of the device and the external environment.

A top view of the arrangement of two cultivation devices 1, each comprising four wells 6 around one central well 7 for the exchange of media, on a common base 11i is shown in Fig. 7.

In a preferred embodiment of the present invention, one cultivation device 1 is arranged on a common base 11 in combination with one in vitro fertilization device 12 as illustrated in Figures 8A-B. In addition, FIG. 8A shows a preferred embodiment of a cultivation device 1 having six wells 6 arranged around a medium change well 7, the longer axes of which run parallel to each other.

The most preferred embodiment of the present invention comprises one removable lid 10 which is common to and which, in the first position, as shown in Fig. 9A, allows access to the internal spaces 5 of said devices 1 and / or 12 arranged on a common base 11, and 9B, in its second, horizontally rotated position relative to the first position, overlaps all the cultivation devices 1 and / or the other one or more devices 12 arranged on a common base 11.

Thus, the specially designed lid shape allows access to the embryos / cells for routine handling, without the lid having to be removed. The lid overlaps the devices 1 and / or 12 arranged on the common base 11, or exposes them, by simply rotating the lid horizontally by 90 °.

Claims (15)

Patent claims A method of adding, removing or replacing a liquid medium in the cultivation and / or manipulation of biological cells, such as γ embryonic cells, deposited in one or more wells embedded in the bottom wall of a culture device, each well directed to its interior with its open upper end the culture device, without any cell manipulation or contact of the cell manipulation device 10, comprising the step of adding media to the culture device, wherein at this step the level of the media in the culture device is gradually raised above the open ends of the wells, thereby forming a fluid communication between the medium to be added and the inner 15 space of said wells through the open ends of the wells, and / or the step of removing the medium from the culture apparatus, wherein the level of the medium in the culture apparatus is gradually decreases below the level of the open ends of said wells, thereby interrupting the connection 20 via the liquid through the open ends of the wells between the added medium and the interior of the wells and the wells will remain inside the space filled with the medium. A method according to claim 1, characterized in that 25 the medium is added to or removed from a well embedded in the bottom wall of the culture apparatus, A cultivation device (1) having a bottom wall (2) and a side wall (3) defining an interior space (5) of the cultivation device (1), 30, characterized in that at least one elongated culture well (6) with a cross-sectional view along its long axis, substantially V-shaped, is recessed in the lower wall (2), with its upper open end facing the interior space (5) of the device (1) ) and at least one well (7) for exchanging the medium, with its upper open end facing the interior (5) of the device (1). Cultivation device according to claim 3, characterized in that the longitudinal walls of the culture well (6) form an angle of 40 to 60 °. Cultivation device according to claim 4, characterized in that the bottom of the culture well (6) is horizontal. Cultivation device according to any one of claims 3 to 5, characterized in that the edge of the upper open end of the culture well (6) facing the interior space (5) of the cultivation device (1) is raised above the upper surface (8) of the bottom wall (2). ) of a cultivation device (1), thereby forming an edge (13) with a horizontal upper end, wherein the edge (13) surrounds the upper open end of the culture well (6). Cultivation device according to any one of claims 3 to 6, characterized in that at least a portion of the upper surface (8) of the bottom wall (2) surrounding any of the medium change wells (7) and comprising one or more culture wells (6) has gradient towards the medium change well (7). Cultivation device according to claim 7, characterized in that the angle β of the top surface (8) towards the medium exchange well (7) is 0.5 to 2 ’. Cultivation device according to any one of claims 3 to 8, characterized in that the medium exchange well (7) has conical side walls with a wider end of the well (7) facing upwards. Cultivation device according to any one of claims 3 to 9, characterized in that the culture wells (6) are arranged in the bottom wall (2) of the culture device (1) with longer axes of the culture wells (6) running parallel to each other. The cultivation device according to any one of claims 3 to 10, characterized in that the bottom wall (2) of the cultivation device (1) comprises identification marks. 15 Dec The cultivation device according to any one of claims 3 to 11, characterized in that the cultivation device (1) further comprises a removable lid (4) overlapping the interior space (5) of the cultivation device (1). 20 May Cultivation device according to any one of claims 3 to 11, characterized in that the cultivation device (1) is arranged on a common base with one or more other devices selected from the group of the cultivation device (1) and the in vitro fertilization device (12). . Cultivation device according to claim 13, characterized in that it comprises a single removable lid (10) which is common to all devices arranged on a common base (11) and in its first position overlaps all devices arranged on a common 30, and in its second, horizontally rotated position relative to the first position, allows access to the interior spaces of the devices arranged on the common base (11). Cultivation device according to any one of claims 3 to 14, 5 ', characterized in that the corner at the point of contact between the upper surface (8) of the bottom wall (2) and the side wall (3) is rounded or beveled into the interior of the cultivation device (8). 1).