Rocking device for cell culturing with wide operating volume range and dual-axis rotation

Abstract

A system for the cultivation of biological entities comprises a support platform configured to accommodate a cultivation chamber, a base, and an agitation mechanism connecting the platform to the base. The agitation mechanism is configured to generate movement of the support platform along two distinct axes, including at least one inclination relative to a horizontal plane. In some embodiments, the system comprises a rotation mechanism allowing 360° rotation and an inclination mechanism enabling large tilt angles in opposite directions. The platform may include a cover and a heating system for thermal control. The system is suitable for use with cultivation chambers of various volumes, ranging from 10 milliliters to 200 liters, enabling expansion of biological entities under controlled conditions using batch, fed-batch, or perfusion strategies.

Description

[0001] P3832PC00 Specs (AOFF)

[0002] ROCKING DEVICE FOR CELL CULTURING WITH WIDE OPERATING VOLUME RANGE AND DUAL-AXIS ROTATION

[0003] This application claims the priority of European patent application EP24193725.9, filed on 8 August 2024, the content of which is incorporated herein by reference.

[0004] FIELD OF THE INVENTION

[0005] The present disclosure relates to a system for the cultivation of biological entities such as bioreactors, which are critical in producing pharmaceuticals, viral vectors, and other biotechnological products. The present disclosure relates in particular to bioreactors with an agitation mechanism with multiple agitation axis.

[0006] BACKGROUND

[0007] Traditional systems for the cultivation of biological entities or bioreactors often have limited volume flexibility, requiring different devices or significant adjustments for varying culture scales. These systems also involve high complexity, mechanical intricacy, and operational costs, with a substantial risk of contamination.

[0008] There is a need in the art for bioreactors and systems that provide a very wide operating volume range from lOmL to 50L or to 200L, while ensuring ideal mixing (e.g., of oxygen, pH, and / or the substrate) and minimal stress on the cells.

[0009] Conventional rocker bioreactors cannot accommodate a wide volume range (10mLto 50Loreven 200L) within the same chamber. Current systems require cultures to be transferred to larger chambers as processes scale up, which involves complex manual handling and increased risk of contamination. Current bio reactor systems are constrained from initiating processes at very low volumes due to the extensive surface area of the cultivation chamber and the substantial liquidgas interface. A larger interface between the culture and the chamber increases the likelihood of biological entities being positioned at the liquid-chamber interface, which induces high shear stress, potentially leading to cellular damage or death. Moreover, an increased interface between the culture and the gas phase results in elevated evaporation rates, altering cultivation conditions by increasing the concentration of certain chemical elements, thereby adversely affecting the culture and possibly causing its demise. P3832PC00 Specs (AOFF)

[0010] The disclosure allows the physical segmentation of the culture and limits the liquid-chamber and liquid-gas interfaces by positioning the system to concentrate the culture in a corner or on the side of the cultivation chamber.

[0011] SUMMARY OF INVENTION

[0012] It is therefore one aspect of the present disclosure to provide a system for the cultivation of biological entities or a bioreactor according to claim 1. The bioreactor is able to use cultivation chamber having a hollow interior space (e.g. closed single use bags) to grow biological entities cultures of volumes ranging, for example from 1 mL to 50 L or 10 mL to 50 L or1 mL to 200L or 10 mL to 200 L or 50 L to 200 L greater. Other advantageous features can be found in the dependent claims.

[0013] According to a first aspect, the present disclosure relates to a system for the cultivation of biological entities, the system comprising a support platform configured to accommodate a cultivation chamber; a base; and an agitation mechanism configured to move the support platform relative to the base, the agitation mechanism comprising a first end connected to the support platform at an attachment point and a second end connected to the base. The agitation mechanism is configured to generate movement of the support platform along two distinct axes of inclination or rotation, including at least one inclination movement of the support platform relative to a horizontal plane on which the system is placed.

[0014] In one embodiment, the agitation mechanism comprises an inclination mechanism configured to tilt the support platform up to 60 degrees or 90 degrees in a first direction and up to 12 degrees or 45 degrees in a second opposite direction relative to a horizontal plane on which the system is placed.

[0015] In one embodiment, the agitation mechanism comprises a rotation mechanism configured to rotate the support platform around a vertical axis extending through or near the attachment point, with a rotational range of up to 360 degrees.

[0016] In one embodiment, both movements generated by the agitation mechanism are inclination movements occurring in different planes.

[0017] In one embodiment, the first end of the agitation mechanism is connected to the support platform at a single attachment point. P3832PC00 Specs (AOFF)

[0018] In one embodiment, the single attachment point is located at the rotation axis.

[0019] In one embodiment, the single attachment point is located in an eccentric position relative to the geometric center of the support platform.

[0020] In one embodiment, the agitation mechanism comprises a first actuator and a second actuator.

[0021] In one embodiment, the first actuator and the second actuator are configured to be actuated independently.

[0022] In one embodiment, the biological entities are selected from the group consisting of human cells, animal cells, insect cells, microbial cells, plant cells and virus.

[0023] In one embodiment, the support platform comprises a coverthat encloses the cultivation chamber.

[0024] In one embodiment, the system comprises a heating system configured to heat the cultivation chamber.

[0025] In one embodiment, the system is configured to support a cultivation chamber comprising a volume ranging from 10 milliliters to 50 liters, or from 10 milliliters to 200 liters.

[0026] It is another aspect to provide a method for expanding biological entities according to claim 14, the method comprising the steps of placing a cultivation chamber on the support platform of the system as defined in any one of the preceding claims, introducing a culture medium and biological entities into the cultivation chamber, and actuating the agitation mechanism to promote cellular expansion.

[0027] In one embodiment, the method comprises carrying out the expansion of biological entities in batch mode, fed-batch mode, perfusion mode, or using a combination of an expansion phase and a production phase.

[0028] Biological entities comprise human cells, animal cells, insect cells, microbial cells, virus, or plant cells.

[0029] The cultivation chamber is mounted on a support platform. P3832PC00 Specs (AOFF)

[0030] The support platform comprises a heating system to maintain the cultivation chamber at the desired temperature.

[0031] Additionally, a mobile heating system can be used to heat specific parts of the cultivation chamber.

[0032] The bioreactor system controls the position of the platform via two rotation axes fixed at a point on the support platform. A first axis can move the platform up to 90° forwards and 45° backwards. A second axis can allow full rotations.

[0033] One important innovation resides in the fact that the system allows the use of an additional 360° rotation axis, enabling orbital and lateral movements which are not possible with a traditional rocker, but also in the fact that the rocker axis allows a much wider rotation ranging from 90° forward to 45° backward, thanks to the shape of the system base and the agitation mechanism.

[0034] The system high degree of freedom of movement means that the culture chamber can be manipulated in any possible configuration that both axes allow. The system also moves the cultivation chamber like a bioreactor rocker device as disclosed in the document US 6,190,913.

[0035] The bioreactor according to the disclosure allows greater control and flexibility in mixing the culture chamber.

[0036] For example, it is possible to concentrate the culture media, such as liquid in a corner of the cultivation chamber, to enable ideal mixing without too much shear stress on the bag surface, especially at small volume.

[0037] The system can be combined with modules to control temperature, pH, oxygen concentration, cell concentration, substrate concentration or any other suitable system to monitor culture parameters in the culture chamber. The innovation comprises a screen and / or a control system connected directly to the equipment or remotely.

[0038] The disclosure also comprises several improved cultivation chambers or bags according to claim 4 designed to be used in the present system.

[0039] The present system enables the cultivation of biological entities under dynamic and controllable conditions by combining dual-axis agitation with precise thermal regulation. The dual-axis movement of the support platform allows more efficient mixing of the culture medium while P3832PC00 Specs (AOFF) reducing shear stress, thereby improving cell viability and uniformity of growth. This is particularly advantageous for sensitive cell types requiring gentle agitation.

[0040] The ability to generate complex agitation profiles through independent inclination or rotation mechanisms improves the homogeneity of nutrient and gas distribution inside the cultivation chamber. Moreover, by enabling directional control of the fluid movement, the system allows targeted concentration of the culture medium or biological entities in specific areas of the cultivation chamber, which can be beneficial in early-stage culture or when transitioning to a production phase.

[0041] The inclusion of a heating system integrated into the platform, the bag support, or the cover ensures uniform and responsive thermal control, facilitating optimal growth conditions for a wide range of biological entities. Thermal precision is further improved by the possibility of integrating temperature sensors and air-flow regulation, enabling adaptive heating depending on the culture stage or medium volume.

[0042] The system is designed to be compatible with different cultivation strategies, including batch, fed- batch, and perfusion modes. This versatility supports applications in biotechnology, cell therapy, vaccine production, and other fields requiring scalable and adaptable cell expansion platforms.

[0043] Furthermore, the modular structure of the system, including a single attachment point and a removable cover, simplifies installation and maintenance of cultivation chambers, while maintaining sterility and functional integrity throughout the process.

[0044] The above and other objects, features, and advantages of the present disclosure and the manner of realizing them will become more apparent, and the disclosure itself will best be understood from a study of the following description with reference to the attached drawings showing some preferred embodiments of the disclosure.

[0045] BRIEF DESCRIPTION OF THE DRAWINGS

[0046] The accompanying drawings, which are incorporated herein and constitute part of this specification, illustrate the presently preferred embodiments of the disclosure, and together with the general description given above and the detailed description given below, serve to explain features of the disclosure. Thicknesses of layers / elements, and sizes of components / elements, are not necessarily drawn to scale or in actual proportion to one another but rather are shown as example representations. Like reference numerals may refer to like parts throughout the several P3832PC00 Specs (AOFF) views. Each embodiment herein may be used in combination with any other embodiment(s) described herein.

[0047] The above and other objects, features, and advantages of the present disclosure and the manner of realizing them will become more apparent, and the disclosure itself will best be understood from a study of the following description with reference to the attached drawings in which:

[0048] Figure 1A, 1 B and 1 C illustrate an embodiment of the system for the cultivation of biological entities, with a flat support platform.

[0049] Figure 2 illustrates an embodiment of the system performing an orbital motion: the corner of the platform may be put in rotation using the coordinated motion of the 2 axis of rotation.

[0050] Figure 3 illustrates a lateral or rotational motion of the system according to an embodiment.

[0051] Figure 4A, 4B and 4C illustrate an embodiment of a rocking motion from +90° to -45° that passes through 0°.

[0052] Figure 5A and 5B illustrate an embodiment comprising a bigger support platform arranged on the support platform.

[0053] Figure 6A illustrates the arrangement of motors, gearboxes, shaft and the structural element that physically fixes them according to an embodiment of the present disclosure. The system is arranged to move the support platform relative to the base.

[0054] Figure 6B illustrates a solution of attaching the motor mechanism, shaft and reducer to the structure according to an embodiment of the present disclosure. It also illustrates how this structure maintains its stability.

[0055] Figure 7 A illustrates a platform with a bag arranged on the platform according to an embodiment of the present disclosure.

[0056] Figure 7B illustrates a platform fitting the shape of the bag and the lid. according to an embodiment of the present disclosure

[0057] Figure 7C illustrates a platform fitting the shape of the bag as well as a representation with the bag and the lid according to an embodiment of the present disclosure. P3832PC00 Specs (AOFF)

[0058] Figure 7D illustrates a larger platform with a bag arranged on the platform according to an embodiment of the present disclosure.

[0059] Figure 7E illustrates a larger platform fitting the shape of the bag and the lid according to an embodiment of the present disclosure.

[0060] Figure 7F illustrates a larger platform fitting the shape of the bag as well as a representation with the bag and the lid according to an embodiment of the present disclosure.

[0061] Figure 8A illustrates the platform positioned in a way that concentrates the liquid in the corner of the cultivation chamber according to an embodiment of the present disclosure.

[0062] Figure 8B illustrates the platform positioned in a way that concentrates the liquid in the side of the cultivation chamber according to an embodiment of the present disclosure.

[0063] Figure 9A illustrates a pillow style two-dimensional bag construction according to the prior art.

[0064] Figure 9B illustrates a pillow style two-dimensional bag construction with two cornerwith a round shape according to an embodiment of the present disclosure.

[0065] Figure 9C illustrates a pillow style two-dimensional bag construction with one side being a halfcircle according to an embodiment of the present disclosure.

[0066] Figure 9D illustrates a pillow style two-dimensional bag construction with four corners with a round shape according to an embodiment of the present disclosure.

[0067] Figure 9E illustrates a pillow style two-dimensional bag construction with two sides being a semiellipse according to an embodiment of the present disclosure.

[0068] Figure 10 illustrates the system with a closed platform according to an embodiment of the present disclosure.

[0069] Figure 11 illustrates the system with a closed platform according to an embodiment of the present disclosure from the front view and the arrow illustrates the lateral motion of the platform. P3832PC00 Specs (AOFF)

[0070] Figure 12 illustrates the system with a closed platform according to an embodiment of the present disclosure from the side view and the arrow illustrates the rocking motion of the platform.

[0071] Figure 13 illustrates the system with a opened platform according to an embodiment of the present disclosure.

[0072] Figure 14 illustrates the heating system and lighting support mounted on the side of the bag according to an embodiment of the present disclosure.

[0073] Figure 15 illustrates heating system integrated into the bag attachment structure according to an embodiment of the present disclosure.

[0074] Figure 16 illustrates heat pads positioned under the bag support platform according to an embodiment of the present disclosure.

[0075] Figure 17 illustrates the position of the observation camera according to an embodiment of the present disclosure.

[0076] Figure 18 illustrates a general view of a mechanism behind the system casing according to an embodiment of the present disclosure.

[0077] Figure 19 illustrates the actuating mechanism that drives the system in rocker mode according to an embodiment of the present disclosure.

[0078] Figure 20 illustrates the actuating mechanism that drives the system in lateral movement mode according to an embodiment of the present disclosure.

[0079] Figure 21 illustrates the mechanical arrangement of the two actuation systems that enable both types of movement according to an embodiment of the present disclosure.

[0080] Figure 22 illustrates the system according to another embodiment with a closed platform from the front view and the arrow illustrates the lateral motion of the platform according to an embodiment of the present disclosure.

[0081] Figure 23 illustrates the system illustrated in Figure 22 with a closed platform from the side view and the arrow illustrates the rocking motion of the platform according to an embodiment of the present disclosure. P3832PC00 Specs (AOFF)

[0082] Figure 24 illustrates the mechanical arrangement of the two actuation systems of the system illustrated in Figures 22 and 23.

[0083] Figure 25 summarizes key results and parameters of an embodiment of the present disclosure in an overview table.

[0084] Figure 26 illustrates data of viable cell density and total cells obtained in a system according to an embodiment of the present disclosure

[0085] Figure 27 illustrates data of volume and viability obtained in a system according to an embodiment of the present disclosure

[0086] REFERENCE NUMERALS IN DRAWINGS

[0087] 100 System for the cultivation of biological entities

[0088] 1 Support platform

[0089] 2 System base

[0090] 3 Mechanism cover

[0091] 4a Cultivation chamber fastening strip for short side of platform

[0092] 4b Cultivation chamber fastening strip for long side of platform

[0093] 5 Fixation means or nuts for holding frame support

[0094] 6 Bigger support platform

[0095] 7 Rotation axis of rotation motor

[0096] 8 Arrow representing the orbital motion of the corner of the platform

[0097] 20 Motor actuating the rocking or inclination motion

[0098] 21 Gearbox actuating the rocking or inclination motion via the rotating shaft

[0099] 22 Rotating shaft

[0100] 23 Motor actuating rotational motion of the support platform

[0101] 24 Gearbox actuating rotational motion of the support platform

[0102] 25 Support frame of the system

[0103] 26 Lower part of the support frame of the system

[0104] 27 Linear rail shaft support

[0105] 40 Cultivating chamber having a hollow interior space (bag)

[0106] 41 Cover for the bag

[0107] 42 Platform support that fits the shape of the bag

[0108] 43 Bigger cultivating chamber having a hollow interior space (bag) P3832PC00 Specs (AOFF)

[0109] 44 Cover for a bigger bag

[0110] 45 Platform support that fits the shape a bigger bag

[0111] 50 Schematic illustration of a culture chamber on the support platform

[0112] 51 Liquid position inside culture chamber in orbital mode

[0113] 52 Liquid position inside culture chamber in lateral mode

[0114] 110 Pillow style two-dimensional bag construction according to the prior art

[0115] 111 Pillow style two-dimensional bag construction with two corners with a round shape

[0116] 112 Pillow style two-dimensional bag construction with a side being a semi-ellipse

[0117] 113 Pillow style two-dimensional bag construction with four corners with a round shape

[0118] 114 Pillow style two-dimensional bag construction with two sides being a semi-ellipse.

[0119] 121 Welding two plastic films together

[0120] 122 Round corner welding

[0121] 123 Semi-elliptical side welding

[0122] 201 System base

[0123] 202 Platform

[0124] 203 Right platform cover

[0125] 204 Left platform cover

[0126] 205 Support leg for base

[0127] 206 Mechanism cover

[0128] 210 Support structure for bag clamp and bag heater

[0129] 211 Bag fastening strip

[0130] 212 Actuating screw for bag clamp.

[0131] 213 Hole through platform for tubing, fiber optics and cables

[0132] 214 Opening cover locking mechanism

[0133] 215 Heating box and camera support element

[0134] 216 Inlet grid of the support structure for bag clamp and bag heater

[0135] 217 Outlet grid of the support structure for bag clamp and bag heater

[0136] 218 Inlet grid of the heating system of the platform

[0137] 219 Heating system of the platform

[0138] 220 Outlet grid of the heating system of the platform

[0139] 221 Heating pads located under the bag support platform.

[0140] 222 Mechanism for raising the cover on the left-hand side for easy opening.

[0141] 223 Platform on which the cultivation chamber may be installed.

[0142] 230 Camera

[0143] 231 Lighting system

[0144] 240 Support structure

[0145] 241 Rocker mode actuator mounting system P3832PC00 Specs (AOFF)

[0146] 242 Rocker mode actuating mechanism motor

[0147] 243 Gearbox for rocker mode actuation mechanism

[0148] 244 Flexible coupling

[0149] 245 Transmission part to lateral mode actuation system

[0150] 246 Lateral mode actuation system motor

[0151] 247 Gearbox for lateral mode actuation mechanism

[0152] 248 Mounting system for coupling gearbox to platform

[0153] 250 Mounting system for coupling gearbox to platform

[0154] 251 Motion transmission component

[0155] 252 Gearbox

[0156] 253 Actuator

[0157] 254 Support structure

[0158] DETAILED DESCRIPTION OF EMBODIMENTS

[0159] The following detailed structural and / or functional description^) is / are provided as examples only, and various alterations and modifications may be made. The example embodiments herein do not limit the disclosure and should be understood to include all changes, equivalents, and replacements within ideas and the technical scope herein. Hereinafter, certain examples will be described in detail with reference to the accompanying drawings. When describing various example embodiments with reference to the accompanying drawings, like reference numerals may refer to like components and a repeated description related thereto may be omitted.

[0160] In a first aspect, the present disclosure concerns a system 100 for the cultivation of biological entities, or a biological entity cultivation system 100.

[0161] The system comprises a platform 1 , 201 and a base 2, 202 (Fig. 1A, 1 B, 1 C, 10).

[0162] The platform may support one or multiple culture chambers or bag that have a hollow interior space 40 43 and that may regulate its temperature.

[0163] The system 100 comprise an agitation mechanism or a dual-axis agitation mechanism. The agitation mechanism comprises a first end connected to the support platform 1 , 201 at an attachment point and a second end connected to the base 2 (Fig. 6A, 6B, 11 , 12).

[0164] The base can contain the mechanism and motors that operate the support platform and / or the agitation mechanism. In addition to this function, the base may also contain a system controller. P3832PC00 Specs (AOFF)

[0165] The base 2, 201 may define an internal compartment configured to house actuation components arranged to operate the support platform 1 , 202, 223 and the agitation mechanism. These components may include one or more actuators, shafts, couplings, or transmission elements. The base may further comprise structural or protective features allowing mechanical integration, thermal management, and service access.

[0166] The (dual-axis) agitation mechanism is configured to generate movement of the support platform 1 , 202, 223 relative to the base 2, 201. The movement may be relative along two distinct axes. These axes may define movements of inclination, of rotation, or a combination thereof.

[0167] At least one of the two movements is an inclination movement of the support platform relative to a horizontal plane on which the system 100 is placed.

[0168] The ability to generate a plurality of agitation movements improves the mixing efficiency and enhances the homogeneous distribution of the biological entities within the culture medium inside the cultivation chamber 40, 43. Moreover, the increased freedom and variability of agitation patterns allow for intentional positioning of the culture medium and / or the biological entities in a specific location of the cultivation chamber, such as in a corner or along a side, which may be particularly advantageous for reducing shear stress or optimizing sampling, oxygenation, or thermal transfer. This capability also enables the cultivation of smaller amounts of biological entities, for example at the beginning of the cultivation process, without requiring a smaller cultivation chamber, thereby allowing scale-up in the same vessel.

[0169] Such movement enables the platform to tilt in order to adjust the position and distribution of the liquid culture within the cultivation chamber 40, 43, thereby enhancing mixing efficiency and reducing shear stress.

[0170] In one embodiment, the agitation mechanism comprises an inclination mechanism configured to tilt the support platform 1 , 202, 223 relative to a horizontal plane on which the system 100 is placed. The inclination mechanism may be configured to tilt the platform up to 90 degrees in a first direction and up to 45 degrees in a second, opposite direction. In some variants, the inclination mechanism may be configured to tilt the platform up to 60 degrees in the first direction and 12 degrees in the second direction, or up to 75 degrees forward and 30 degrees backward. In other examples, the inclination range may be symmetrical, for instance 45 degrees in both directions. These values may be selected depending on the application, the size and shape of the cultivation chamber 40, 43, and the desired mixing dynamics. P3832PC00 Specs (AOFF)

[0171] In one embodiment, the agitation mechanism comprises a rotation mechanism configured to rotate the support platform 1 , 202, 223 around a vertical axis 7 extending through or near the attachment point connecting the platform to the agitation mechanism. The rotation mechanism may be configured to allow continuous or partial rotation of the platform, with a rotational range of up to 360 degrees. In some examples, the platform may perform oscillatory movements within a limited angular range, such as ±45 degrees or ±90 degrees, or may rotate continuously in one or both directions. This rotation enables additional mixing patterns and flow dynamics within the cultivation chamber 40, 43, contributing to enhanced distribution of biological entities and dissolved gases throughout the culture medium.

[0172] In one embodiment, the agitation mechanism is configured such that both movements applied to the support platform 1 , 202, 223 are inclination movements occurring in different planes. The two inclination movements may be produced by independent actuators or mechanical linkages, each configured to tilt the platform along a distinct tilt axis. These tilt axes may be oriented orthogonally or at an angle relative to one another, thereby enabling the platform to perform complex compound motions. Such a configuration provides enhanced control over the mixing dynamics inside the cultivation chamber 40, 43, offering improved homogenization of the culture medium and reduced shear stress on the biological entities.

[0173] In one embodiment, the first end of the agitation mechanism is connected to the support platform 1 , 202, 223 at a single attachment point. This single-point connection may serve as a pivot or articulation center for the various movements of the platform. Such a configuration simplifies the mechanical design and facilitates a wider range of compound or coordinated motions by allowing the platform to move freely around the defined attachment point. The term single attachment point refers to a localized structural connection or interface through which the support platform is mechanically linked to the agitation mechanism, such that all movements of the platform are transmitted through this single physical point. This arrangement can also improve the precision and fluidity of the agitation movements, contributing to more effective and uniform mixing of the culture medium within the cultivation chamber 40, 43.

[0174] In one embodiment, the single attachment point connecting the agitation mechanism to the support platform 1 , 202, 223 is located at the rotation axis 7.

[0175] In one embodiment, the single attachment point is located in an eccentric position relative to the geometric center of the support platform 1 , 202, 223. By placing the attachment point off-center, the resulting movement of the platform during rotation or inclination creates more complex trajectories of the cultivation chamber. This eccentric configuration enables specific mixing P3832PC00 Specs (AOFF) patterns and the ability to direct the culture medium or biological entities toward a selected region of the chamber, such as a corner or side, enhancing the adaptability of the system to various culture volumes and conditions.

[0176] In one embodiment, the agitation mechanism comprises a first actuator 20 and a second actuator 23. The first actuator 20 may be configured to generate an inclination movement of the support platform 1 , 202, 223, while the second actuator 23 may be configured to generate a rotation or a second inclination movement, depending on the configuration of the system. The use of two separate actuators provides precise and reliable control of each movement axis, facilitating a wide variety of motion profiles for optimal cultivation conditions.

[0177] In one embodiment, the first actuator 20 and the second actuator23 are configured to be actuated independently. This independent actuation allows the system to perform complex sequences or combinations of movements, such as alternating or simultaneous tilting and rotation, without mechanical interference.

[0178] In one embodiment, the support platform 1 , 202, 223 may be closed using a cover 203, 204, as illustrated in Figures 10 and 13. The cover may be configured to enclose the cultivation chamber positioned on the platform, thereby providing additional protection against contamination and facilitating control of environmental parameters such as temperature, humidity, or gas composition within the enclosed volume. The cover may be detachable or hinged, and may comprise one or more access ports or transparent sections for observation or sampling.

[0179] In one embodiment, the platform 1 , 202, 223 may be secured using a magnetic mechanism 214. The magnetic mechanism may provide a reliable yet easily releasable connection between the platform and associated structural elements or accessories, such as the cover 203, 204. Additionally, a spring mechanism 222 may be fitted to the platform or coverto lift the cover slightly when released, thereby facilitating manual opening. These features, illustrated in Figures 13 and 17, contribute to improved ergonomics and operational convenience during the handling of the cultivation system.

[0180] In one embodiment, the system 100 comprises a cultivation chamber 40, 43 configured to contain a culture medium and biological entities. The cultivation chamber may be implemented as a flexible or semi-rigid bag having a hollow interior space suitable for the growth of cellular entities in suspension or on microcarriers. The cultivation chamber may be removably arranged on the support platform 1 , 202, 223 and may be designed to accommodate a wide range of volumes. P3832PC00 Specs (AOFF)

[0181] The chamber may comprise one or more ports for introducing or extracting fluids, gases, or biological samples, and may be adapted to be sealed or covered using a dedicated lid 203, 204. In some embodiments, the chamber may be shaped to optimize fluid dynamics during agitation, such as having rounded corners or semi-elliptical sides, thereby reducing shear stress and promoting uniform mixing.

[0182] The cultivation chamber or culture chamber 40 43 may be secured to the platform using a fastening system that holds it firmly in place. The fastening system can comprise a strap as disclosed in US 6,190,913 a quick fastener as disclosed in W00066706A1 , a system with fixation means such as nuts 5 212 and a fastening strip 4a 4b 211 (Fig. 1A, 1 B, 13, 15), or other attachment mechanisms.

[0183] In one embodiment, the system is designed to accommodate cultivation chambers with a wide range of working volumes, for example from 10 milliliters to 50 liters, or up to 200 liters. This extended scalability enables the user to perform biological entity expansion across multiple phases using a single system, without the need to transfer the culture between several cultivation chambers of increasing size. As a result, the risk of contamination and handling errors is reduced, and the process is simplified and accelerated.

[0184] In another embodiment, the cultivation chamber comprises a volume ranging from 10 milliliters to 50 liters, or up to 200 liters.

[0185] The platform may contain a heating system which can be controlled by a temperature sensor to maintain a determined temperature in the cultivation chamber.

[0186] To control the temperature of a very specific area of the culture chamber, it is possible to connect a mobile heating system that can be applied to the area of interest. To provide the most precise heating possible, and to avoid heating irrelevant areas, several heating systems may be installed on the platform, and depending on how the platform is used, one heating system or more may be used.

[0187] In one embodiment, the system 100 comprises a heating system 219 configured to heat the cultivation chamber 40, 43 or the contents thereof. The heating system 219 may be integrated into the support platform 1 , 202, 223 or arranged in thermal contact with it, allowing indirect heat transfer to the cultivation chamber. P3832PC00 Specs (AOFF)

[0188] The heating system may comprise one or more electrical resistive elements, flexible heating films, or circulating fluid channels. Temperature regulation may be achieved through feedback control using temperature sensors in proximity to or in contact with the cultivation chamber. This configuration enables precise thermal control of the culture environment, which is critical for the viability and growth of the biological entities.

[0189] In one embodiment, the system 100 comprises multiple heating embodiments to ensure accurate thermal regulation of the cultivation chamber 40, 43. To heat the support platform 223 on which the cultivation chamber is installed, one or more heating mats may be placed beneath a heating plate 221 , as illustrated in Figure 16. In addition, a heating box 215 may be positioned on the support platform 223. The heating box may define a hollow internal volume in which a heating system 219 is arranged. The heating system 219 may comprise one or more electrical resistors and, optionally, one or more fans. The heating box may further include an air inlet 218 and an outlet grid 220 to allow controlled airflow through the heated chamber.

[0190] In another embodiment, a heating system may be integrated within the support structure associated with the bag clamp 210. As illustrated in Figure 15, the support structure may include two lateral suction grids 216 through which ambient air is drawn. The air may be heated internally by the heating system, then directed through internal ducts to the area beneath the cultivation bag. A plurality of holes 217 distributed within the support structure enables uniform diffusion of the heated air across the surface of the bag. This embodiment ensures a precise and homogeneous thermal environment for the cultivation chamber, enhancing cell viability and culture performance even in small-volume or thermally sensitive processes.

[0191] The heating system may be controlled by one or more sensors, depending on how much the culture chamber is filled. The sensors or the temperature sensors configured to monitor the thermal conditions of the cultivation chamber 40, 43 or its surrounding environment. The number and placement of sensors may be selected based on the fill level of the cultivation chamber, allowing the system to adapt the heating power and distribution accordingly.

[0192] In one embodiment, the system may comprise a cooling system. The cooling system may be configured to counteract heat generated by the motors or other actuation components located beneath the support platform 1 , 202, 223. In addition to mitigating unwanted thermal accumulation, the cooling system may also serve to actively lower the temperature of the cultivation chamber 40, 43 when required by the biological process. This allows for precise thermal management across a wide range of cultivation conditions, including processes involving temperature reduction phases or low-temperature maintenance P3832PC00 Specs (AOFF)

[0193] The same principle applies to temperature sensors. One or more can be connected, and depending on the mode of use, either one or a combination of temperature sensors can be used.

[0194] Alternatively, the temperature of the cell culture fluid can be controlled by placing the culture chamber on the platform in a temperature-controlled chamber.

[0195] The platform may support three primary types of motion: rocking or inclination (Fig. 4A, 4B, 4C, 12, 23), lateral or rotational (Fig. 3, 11 , 22), and orbital (Fig. 2). This versatility is achieved through the agitation mechanism or the dual-axis agitation mechanism having a first end connected to the support platform at an attachment point and a second end connected to a base and configured to generate all types of motion, individually or in combination.

[0196] The dual-axis agitation mechanism may comprise two actuators or motors 20, 23, 242, 246 capable of generating all movement types, individually or in combination.

[0197] These two actuators or motors may be part of the system base.

[0198] This base may comprises of a support frame 25 240 that allows the entire system to be placed on the floor or on a table. This support frame may comprises of a lower part 26 which ensures that the system is stable and remains in equilibrium (Fig. 6B).

[0199] The motor or actuator or actuation 20242 that powers or actuate the inclination or rocking motion may be attached to the system support frame 25 240. This motor is referred to as the rocking motor 20 242. The rocking motor may be directly coupled and fixed to a gearbox 21 243 that allows speed and torque to be adapted. This gearbox may be connected to a rotating shaft 22 that can be set in motion. This example of motorization system enables the system to perform a rocking movement.

[0200] The second motorization or actuation 23 246 may fixed to the shaft 22 or a support frame 245 that can be actuated by the rocking motor 20 242. This motor, known as the rotation motor, may drive the rotation of the platform. The shaft 22 may be attached to the rotation motor gearbox 24 via a linear rail shaft support 27. The rotation motor may be directly attached and coupled to this gearbox 24 247. This gearbox transforms the speed and force applied by the motor to the needs of the system. The gearbox 24 247 may be attached to the cultivation chamber support platform via a support plate 28. P3832PC00 Specs (AOFF)

[0201] The system may comprise an inclination motion and / or a rocking motion. In this case, the movement of the rocking motor 20 generates a movement like those described in US 6,190,913 and W00066706A1 , but the present system allows a larger magnitude of movement from +90° to -45° or +60° to -12, or from +75° to -30°, or from +45° to -15°, depending on the specific cultivation requirements. These different tilt ranges may be selected based on the viscosity of the culture medium, the sensitivity of the biological entities, orthe desired mixing profile. This flexibility provides enhanced adaptability to a variety of cell culture protocols and cultivation chambers geometries. (Fig. 4A, 4C, 23). The rocking movement is represented by the arrow in Figure 4B and Figure 23.

[0202] The 0° position (Fig. 4B) corresponds to the state where the plane of the support platform is parallel to the ground plane, or in Figure 4B to the y axis. The +90° position corresponds to the state where the platform plane is positioned at +90° or perpendicular to the ground or in Figure 4Ato the y axis. The -45° position corresponds to the state where the platform plane is positioned at -45° to the ground or in Figure 4C to the y axis.

[0203] Lateral movement, represented by the arrow in the Figure 3 and Figure 11 , may be driven by the movement of the rotation motor 23, 246.

[0204] To perform the lateral movement, the platform may be positioned at around +90° to the ground by the rocking motor as represented in Figure 4A, and then the rotation motor may be operated exclusively to perform movements around the rotational axis 7 shown in the Figure 3.

[0205] In this case, the platform may be positioned in a way that concentrates the liquid 52 in the side of the cultivation chamber (Fig. 8B). The platform may be placed vertically compared to the ground. The lateral movement may generate liquid motion within the culture chamber on the side of the cultivation chamber (Fig. 3 and 8B). Total rotation may be possible with this motor.

[0206] Orbital movement (Fig. 2) may be produced by synchronized actuation of the two motors, resulting in a motion like an orbital shake. In this configuration, the platform may move in a circular path parallel to the ground 8 or parallel the plane xy in the Figure 2, with the corner point of the platform maintaining a consistent radius from a central fixed point. This central point of rotation may remain stationary relative to the ground, ensuring that the corner of the platform orbits around it. It may also be possible to make elliptical movements that draw an ellipse in relation to the central point of the platform corner. When performing the orbital movement the liquid is concentrated in the corner 51 as shown in the Figure 8A, and the movement mixes the cell culture like an orbital shaker. P3832PC00 Specs (AOFF)

[0207] These are three types of movement in example, but the system is not limited to these three types of movement alone. It may be possible to combine the two-motion mechanism, inclination and rotation, and make all possible movements within the physical limits allowed by the two axis. The system may also enable the simultaneous combination of lateral and rocking movements. The system may operate for both axis at frequencies ranging from about 0 to 40 cycles per minute or greater.

[0208] Figures 22 and 23 illustrate an embodiment of the device comprising a first rocker axis configured to operate up to maximum angles between -15° and 90° (as indicated by the arrow in Figure 23), and a second rocker-type axis arranged along an axis perpendicular to the first rocker axis (as indicated by the arrow in Figure 22), the second rocker-type axis being configured to operate up to maximum angles between +25° and -25°. The system can be positioned at any of these angles, and it is possible to actuate either one of the movements individually or both movements in combination.

[0209] A key advantage of the system according to the disclosure over existing bioreactor rocking systems is its ability to precisely control mixing within the culture chamber. The system can position the culture liquid in various locations, such as a corner 51 , one side 52, or along the entire length of the cultivation bag 40.

[0210] Control on two axes offers numerous combinations for improved mixing, enhancing the regulation of elements such as pH, carbon dioxide and oxygen concentration by enabling faster and more uniform distribution of chemical compounds in the liquid.

[0211] According to one aspect of the disclosure, the system is configured to vary the rocking angle to a maximum of 90 degrees along a first axis and to actuate a second rocker-type axis arranged perpendicular to the first axis. The system is further configured to modulate the contact surface area between the solid phase of the culture chamber and the culture medium by adjusting the angle of rotation along one or both axes.

[0212] Such modulation enables control of the shear stress exerted on the culture medium as well as the thickness of the culture medium film, thereby optimizing gas exchange processes. Furthermore, the system permits adjustment of the contact surface between the gaseous phase within the culture chamber and the culture medium, thereby improving gas transfer efficiency.

[0213] By combining these physical parameters, namely modulation of the contact surface between the culture medium and the solid phase and / or the gaseous phase, the system provides enhanced P3832PC00 Specs (AOFF) flexibility to optimize cell culture conditions. This is achieved by precisely tuning mixing dynamics, shear stress, film thickness, and gas exchange in accordance with the specific requirements of the cell culture process.

[0214] The present disclosure also comprises several improved cultivation chambers designed to be used in the present system (Fig. 9B, 9C, 9D, 9E).

[0215] Figure 9A illustrates the construction of a two-dimensional pillow-type bag 110 according to the prior art (US 6,190,913). The shape of this cultivation chamber may work with the innovation presented, but the design of the corners of this construction is not always optimal for the type of cell, e.g. cells sensitive to shear stress may be impacted by this design. Particularly in the case of lateral or orbital movement, having a bag with square corners is not optimal in terms of mixing, as the liquid at the tip of the corner is more difficult to mix, and the movement generated in this configuration causes more shear stress.

[0216] For this reason, variants of the bag shown in Figure 9A has been invented.

[0217] The aim of these new bags is to limit cell shear stress, and this is achieved by having round corners.

[0218] Four embodiments of the bag are shown in Figures 9B, 9C, 9D and 9E.

[0219] The bag 111 in figure 9B comprises two rounded corners 122.

[0220] The bag 112 in figure 9C comprises one side being a semi-ellipse 123.

[0221] The bag 113 in figure 9D comprises four rounded corners 122.

[0222] The bag 114 in figure 9D comprises two sides being a semi-ellipse 123.

[0223] The embodiments depicted in Figure 9B and 9C exhibit a configuration wherein one side is rounded (either by having two round corners 122, or by having one semi-elliptical side 123), and the opposing corners maintain a rectangular form akin to a standard bag 110.

[0224] This design is formulated to enhance mixing efficiency and mitigate shear stress when the system is operated in orbital or lateral mode, owing to the round shape. P3832PC00 Specs (AOFF)

[0225] The bag may be oriented with the rounded side or corners positioned downward, thereby concentrating the liquid in the corner or on the rounded side during orbital or lateral mode operation. The rectangular form on the opposite side is intended to optimize mixing in rocker mode.

[0226] Figures 9D and 9E illustrate embodiments of the bag comprising rounded edges (either by having two round corners 122, or by having one semi-elliptical side 123) on two opposite sides. This configuration, like the previous designs, improves mixing and minimizes shear stress in orbital or lateral mode. Additionally, the rounded edges on the two opposing sides serve to diminish shear stress in rocker mode.

[0227] These bags may be obtained by welding 121 two plastic films together. The bags may be equipped with or comprise multiple ports to allow gas, liquids, cells, etc. in or out.

[0228] The cultivation chamber 40, 43 may be implemented as a flexible bag formed by welding 121 two plastic films together. The bag may define a sealed internal volume suitable for containing a culture medium and biological entities. The bag may comprise one or more functional features, such as ports configured for the introduction or extraction of gases, liquids, cells, or sampling probes. These ports may include sterile connectors, filters, or valves, depending on the process requirements. The bag may also be designed to maintain sterility and structural integrity during agitation, heating, and cooling, and may optionally include reinforced zones, gripping areas, or markings for alignment with the support platform or clamping mechanisms.

[0229] The bag can be clamped in such a way as to leave a section empty of gas or liquid, thereby creating at least one section or several sections intentionally left free of gas or liquid.

[0230] This disclosure is suitable for culturing a wide range of biological entities, comprising human cells, animal cells, plant cells, microbial cells, and / or insect cells, in both suspension and anchoragedependent systems. It is also effective for cultivating viruses and pathogens.

[0231] The system according to the disclosure may also comprise components such as a control unit, speed control, heater, tachometer, and sensors for monitoring parameters like pH, oxygen, carbon dioxide, cell concentration, nutrients, temperature in the cell culture chamber, humidity, pressure, fluid levels, and more, ensuring comprehensive control and optimization.

[0232] The system may also comprise means that may provide automatic control of pH, dissolved oxygen (DO), and capacitance, enabling precise regulation of the culture environment. These P3832PC00 Specs (AOFF) means may include sensors for real-time monitoring, actuators that can adjust gas composition, liquid additions or removals, and software or algorithms that may dynamically adjust process parameters. The pH may be controlled by injection of gases such as carbon dioxide, by addition of acid or base solutions, or by any other element or method that may influence the pH. The DO may be controlled by modifying the oxygen concentration in the gas mixture, by adjusting the pressure, by increasing or modifying the speed or angle of mixing, or by any other element or method that may affect the dissolved oxygen level. The capacitance measurement may allow the system to maintain the cell culture at a specific viable cell density by controlling a pump connected to a medium reservoir, enabling automated feeding, dilution, perfusion, bleeding or any other action that may contribute to maintaining the desired cell density and culture conditions.

[0233] The process carried out in the system may include cell expansion or production using various modes of operation, such as batch, fed-batch, or any combination of expansion and production phases. The system may also be configured to perform perfusion processes, which can involve continuous or bolus addition of fresh culture medium, removal of supernatant, and cell bleeding or any other action that may support steady-state or intensified culture conditions. These modes of operation may be applied independently or in combination, depending on the desired process strategy.

[0234] Perfusion may be implemented using a bag with an integrated perfusion device, such as an alternating tangential flow (ATF) system, a tangential flow filtration (TFF) system, or any other existing or future technology that may enable perfusion of the culture.

[0235] In an embodiment, the system is capable of handling for example a cultivation chamber comprising a volume up to 30 L of liquid culture volume and has overall dimensions of approximately 120 cm in width, 80 cm in depth, and 110 cm in height. These compact dimensions make the system suitable for use in standard laboratory environments while still enabling significant culture volume scalability within a single-use format.

[0236] In use, the present system may be employed for the cultivation, amplification, or production of a wide variety of biological entities, including but not limited to mammalian cells, microbial cells, insect cells, plant cells, or viral particles. The system is particularly suited for applications in biotechnology, biopharmaceutical manufacturing, and cell therapy, where precise control of environmental conditions and gentle agitation are essential to maintain cell viability and functionality. For example, the system may be used for the expansion of CHO cells for monoclonal antibody production, the amplification of T cells or NK cells for adoptive immunotherapy, the culture of HEK293 cells for viral vector production, or the growth of engineered bacteria for P3832PC00 Specs (AOFF) recombinant protein expression. The system may also be used in research or clinical settings to produce exosomes, cultivate stem cells, or maintain organoid cultures under controlled and reproducible conditions. Its flexibility in agitation and thermal control makes it suitable for both small-scale development and large-scale manufacturing processes.

[0237] Method for Cultivating Cells in a Flexible Bag Bioreactor or a cultivation chamber

[0238] The present disclosure also concerns a method for expanding biological entities or a method for cultivating biological entities.

[0239] In one embodiment, a method for expanding biological entities comprises the steps of: placing a cultivation chamber 40, 43 on the support platform 1 , 202, 223 of the system 100; introducing a culture medium and biological entities into the cultivation chamber; and actuating the agitation mechanism to promote cellular expansion under controlled conditions. The cultivation chamber may be in the form of a flexible or semi-rigid bag and may be secured to the platform using dedicated clamps or holders. The agitation mechanism may then be activated to move the platform along two distinct axes, thereby enhancing the homogeneity of mixing, ensuring uniform distribution of nutrients, and minimizing sedimentation of the cells. The system allows for fine control over agitation intensity and duration, which may be adapted to the sensitivity and growth phase of the biological entities. Environmental parameters such as temperature may also be regulated during the expansion process using the integrated heating or cooling systems. This method enables the reproducible expansion of various biological entities including mammalian cells, microbial cells, plant cells, insect cells, or virus-producing cell lines, in research, development, or manufacturing settings.

[0240] In one embodiment, the expansion of biological entities using the present system may be carried out in batch mode, fed-batch mode, or perfusion mode. In batch mode, all components required for cultivation are introduced into the cultivation chamber at the beginning of the process, and no additional input is provided until harvest. In fed-batch mode, nutrients or other supplements may be gradually added during cultivation to sustain growth over a longer period and improve yield. In perfusion mode, the culture medium is continuously or intermittently refreshed while retaining the biological entities within the cultivation chamber, enabling prolonged cultivation and higher cell densities. The system may also be used to perform a combination of an expansion phase followed by a production phase, for example in the context of biopharmaceutical manufacturing where cells are first amplified under optimal growth conditions and then stimulated to produce a target compound. The flexibility of the agitation and control mechanisms of the system supports adaptation to these various modes of operation. P3832PC00 Specs (AOFF)

[0241] In another embodiment, the method for cultivating cells may comprise one or several of the following steps or some of the following steps:

[0242] Placing a flexible cultivation bag on the platform of the bioreactor system;

[0243] Introducing a sterile gas mixture into the bag to maintain the necessary gaseous environment for cell culture;

[0244] Filling the bag with a culture inoculum under sterile conditions; continuously monitoring and regulating the temperature of the bag throughout the process to ensure optimal culture conditions;

[0245] As the cell culture multiplies, incrementally increasing the medium volume to maintain optimal cell concentration, achieved either through manual addition of medium or by automated pumping;

[0246] When the initial cell culture volume is low, activating the orbital operation mode of the bioreactor to gently mix the culture;

[0247] As the culture volume increases, transitioning the system to lateral agitation mode to ensure proper mixing and aeration; and upon reaching a sufficient culture volume, switching the system to rocking mode to maintain effective mixing and aeration, minimizing shear stress and ensuring homogeneous conditions throughout the culture.

[0248] This method ensures controlled and efficient scaling of cell cultures within a flexible bag bioreactor, optimizing cell growth and minimizing potential issues related to gas exchange and shear stress.

[0249] While the disclosure has been disclosed with reference to certain preferred embodiments, numerous modifications, alterations, and changes to the described embodiments, and equivalents thereof, are possible without departing from the sphere and scope of the disclosure. Accordingly, it is intended that the disclosure not to be limited to the described embodiments and be given the broadest reasonable interpretation in accordance with the language of the appended claims. The features of any one of the above-described embodiments may be included in any other embodiment described herein.

[0250] Example of Cell Culture Process P3832PC00 Specs (AOFF)

[0251] The following examples are provided for illustrative purposes only and are not intended to limit the scope of the present disclosure.

[0252] In one example, Chinese Hamster Ovary (CHO) cells were cultured in a disposable cell culture bag. The cell culture bag was composed of polyethylene (PE) and had a total volume of 60 liters. The bag was inflated with a gas mixture consisting of approximately 95% air and 5% carbon dioxide (CO2). The temperature of the system was maintained at approximately 37°C throughout the entire process.

[0253] The CHO cells were initially thawed in 30 mL of fresh culture medium (powerCHO, Lonza) within a 50 mL centrifuge tube. Following thawing, the cell suspension was transferred to the cell culture bag using a transfer cap equipped with weldable tubing, which was welded directly onto the bag. The cell expansion was initiated with a starting volume of approximately 30 mL.

[0254] The culture volume was increased daily, approximately doubling each day, while maintaining a viable cell density below 2 x 106cells / mL until the culture reached 30 liters. From the initial volume of 30 mL up to approximately 30 liters, the culture was operated in swing mode agitation (from day 0 to day 7). Starting on day 7, the agitation mode was changed to a rocking motion. The culture volume was progressively increased by daily addition of fresh medium until reaching a final volume of approximately 30 liters. The medium was added at a substantially linear rate using a peristaltic pump, with the addition rate corresponding to the culture volume (e.g., approximately 30 mL of medium added between day 0 and day 1 , approximately 60 mL between day 1 and day 2, and so forth).

[0255] Results

[0256] Using the setup described herein, the system enabled consistent and scalable cell cultivation within a single vessel, ranging from an initial volume of 30 mL up to 30 L. Starting from an inoculation of 30 mL at a concentration of 0.32 x i o6cells / mL, the culture was successfully expanded to 30 L, reaching 1 .99 x 1 o6cells / mL within ten days, and further to 9.56 x 1 o6cells / mL after fourteen days (see FIG. 25, FIG. 26, FIG. 27). Throughout the experiment, cell viability was maintained above 90% (see FIG. 27). The observed growth plateaued due to limitations inherent to the specific cell line and culture medium, which typically do not support cell densities exceeding 10 x 106cells / mL.

[0257] The angle, speed, and operating mode parameters are detailed in the table of FIG. 25. Initially, the system operated in a swing / lateral mode. Once the working volume reached approximately P3832PC00 Specs (AOFF)

[0258] 3.5 L, the system was switched to a rocker mode. In this configuration, both the agitation speed and rocking angle were increased to enhance oxygen transfer to the cells. Oxygenation was monitored via a dissolved oxygen (DO) probe integrated into the bag (data not shown).

[0259] An exponential growth phase was observed and maintained until day 11 (see FIG. 26).

Claims

P3832PC00 Specs (AOFF)CLAIMS1. A system (100) forthe cultivation of biological entities, the system comprising: a support platform (1 , 202, 223) configured to accommodate a cultivation chamber (40, 43); a base (2, 201); and an agitation mechanism configured to move the support platform (1 , 202, 223) relative to the base (2, 201), the agitation mechanism comprising: a first end connected to the support platform (1 , 202, 223) at an attachment point; and a second end connected to the base (2, 201); wherein the agitation mechanism is configured to generate movement of the support platform (1 , 202, 223) along two distinct axes of inclination or rotation, including at least one inclination movement of the support platform (1 , 202, 223) relative to a horizontal plane on which the system (100) is placed.

2. The system (100) according to claim 1 , wherein the agitation mechanism comprises an inclination mechanism configured to tilt the support platform (1 , 202, 223) up to 60 degrees or 90 degrees in a first direction and up to 12 degrees or 45 degrees in a second opposite direction relative to a horizontal plane on which the system (100) is placed.

3. The system according to claim 1 or 2, wherein the agitation mechanism comprises a rotation mechanism configured to rotate the support platform (1 , 202, 223) around a vertical axis (7) extending through or near said attachment point, with a rotational range of up to 360 degrees.

4. The system according to any one of claims 1 to 3, wherein both movements generated by the agitation mechanism are inclination movements occurring in different planes.

5. The system according to any one of the preceding claims, wherein the first end of the agitation mechanism is connected to the support platform (1 , 202, 223) at a single attachment point.

6. The system according to the preceding claim, wherein the single attachment point is located at the rotation axis (7).P3832PC00 Specs (AOFF)7. The system according to the preceding claim, wherein the single attachment point is located in an eccentric position relative to the geometric center of the support platform (1 , 202, 223).

8. The system according to any one of the preceding claims, wherein the agitation mechanism comprises a first actuator (20) and a second actuator (23).

9. The system according to the preceding claim, wherein the first actuator (20) and the second actuator (23) are configured to be actuated independently.

10. The system according to any one of the preceding claims, wherein the biological entities are selected from the group consisting of human cells, animal cells, insect cells, microbial cells, plant cells and viruses.11 . The system according to any one of the preceding claims, wherein the support platform (1 , 202, 223) comprises a cover (203, 204) that encloses the cultivation chamber (40, 43).

12. The system according to any one of the preceding claims, wherein the system (100) comprises a heating system (219) configured to heat the cultivation chamber (40, 43).

13. The system according to any one of the preceding claims, wherein the system is configured to support a cultivation chamber (40, 43) comprising a volume ranging from 10 milliliters to 50 liters, or from 10 milliliters to 200 liters.

14. A method for expanding biological entities, comprising: placing a cultivation chamber (40, 43) on the support platform (1 , 202, 223) of the system (100) according to any one of the preceding claims; introducing a culture medium and biological entities into the cultivation chamber; and actuating the agitation mechanism to promote cellular expansion.

15. The method according to claim 14, wherein the expansion if biological entities is carried out in batch mode, fed-batch mode, perfusion mode, or using a combination of an expansion phase and a production phase.

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