MXPA99002775A - Cell/tissue culturing device and method - Google Patents

Abstract

A disposable device and method for axenically culturing and harvesting cells and/or tissue in consecutive cycles. The device consists of a sterilisable transparent and/or translucent disposable container which may be at least partially filled with a suitable sterile biological cell and/or tissue culture medium and/or axenic inoculant and/or sterile air and/or required other sterile additives. The container has means for removing excess air and/or waste gases therefrom, and means for introducing the inoculant and/or culture medium and/or additives therein. The device is characterised by having a reusable harvesting means for enabling harvesting of at least a portion of the medium containing cells and/or tissue when desired, thereby enabling the device to be used continuously for at least one subsequent consecutive culturing/harvesting cycle. The portion of medium containing cells and/or tissue remaining from a previously harvested cycle may serve as inoculant for a next culture and harvest cycle, culture medium and/or additives being provided. The device may thus be used continuously in consecutive cycles, and may be disposed of when it becomes contaminated. In a second aspect of the invention, a battery of these devices, suitably interconnected, enables the scale of production of cells/tissues to be adjusted when required.

Description

DEVICE AND METHOD FOR CULTIVATING CELLS / TISSUES ii '' FIELD OF THE INVENTION The present invention relates to devices for cultivating and axenically harvesting cells and / or tissues, including bioreactors and fermentors. In particular, this invention relates to such devices which are removable but which, however, can be used continuously, for a plurality of consecutive cultivation / harvesting cycles before the elimination of the same. This invention also relates to the groups of such a large and They have been in the matter. The perspective of using such culture techniques economically is for the extraction of metabolites. secondary agents such as pharmaceutically active compounds, various substances to be used in cosmetics, hormones, enzymes, proteins, antigens, food additives and natural pesticides, to be harvested from cultured cells or tissues. Although potentially lucrative, however, this perspective has not been effectively crystallized with bioreactors on an industrial scale, which use animal and plant crops that "grow slowly, due to the high financial costs involved." The technology of the technique For the production of cell and / or tissue cultures on an industrial scale, to be used for the production of such materials, it is based on vitreous bioreactors and stainless steel bioreactors, which are expensive main parts. industrial systems include costly and complicated mixing technologies, such as »expensive and complicated sterile-powered impellers, and some costly fermentors comprise a multi-part air-emulsion construction.The successful operation of these bioreactors often requires the implementation of aeration technologies, which they constantly need to improve. In addition, such bioreactors are sized according to the maximum volume capacity that is required over time. In this way, problems arise when the proportion of the fermenters in the pilot plant is extended to the large-scale fermenters or when the need arises to increase the production beyond the capacity of the existing bioreactors. The alternative for a large capacity bioreactor, mainly to provide a number of bioreactors j! stainless steel or smaller vitreous, whose total volume capacity is adapted to the requirements, while offering a degree of flexibility to increase or reduce the total capacity, however it is much more expensive than the provision of a larger bioreactor. -nico In addition, the implementation costs associated with most stainless steel and glass bioreactors are also high, due to the lower lower productions coupled with the need to sterilize the bioreactors I after each growing cycle. Consequently, the products extracted from the cells or tissues growing in such bioreactors are costly and can not compete commercially at present with comparable products produced with alternative techniques. In fact, only one Japanese company is known that commercially uses the above-mentioned cell / tissue culture technique, I using stainless steel bioreactors. This company i. produces Shikonin, a compound that is used almost exclusively? in Japan. Bioreactor devices on an industrial scale and even on a large scale are traditionally permanent or "semi-permanent" components and neither a description nor a suggestion of the concept of an eli- minaable bioreactor device are known to solve the aforementioned problems, considering the production of large-scale cell / tissue culture, on the other hand, bioreactor and fermenting devices are well known and are directed exclusively at very small-scale production volumes such as, in local fermentation and for laboratory work. These bioreactor devices generally comprise a disposable bag, which typically opens with a cut in order to collect the cell / tissue production, thus destroying any additional utility of the bag. A known, removable bioreactor is produced by Osmotec, Israel, (Agritech Israel, Publication No. 11, Fall 1997, page 19) for small-scale use, such as in laboratory research. This bioreactor comprises a conical bag having an entrance hole, through which the culture medium, air, can be introduced. inoculant and other optional additives and has a volume of only about 1.5 liters. Aeration is carried out by introducing very small air bubbles, which in many cases result in damage to the cells, particularly in the case of plant cell cultures. In particular, these bags are designed specifically for only a single crop / harvest cycle and the contents of the bag are removed i, by cutting the bottom of the bag. Therefore, these bags are not directed towards an economic solution to the problem of providing industrial quantities of the materials to be extracted from the crop, as discussed above. The elimination of these bioreactor devices generally does not present an economic disadvantage to the user since even the lower financial costs of these parts are offset against ease of use, storage and other practical considerations. Of fact, at the levels of production lower than these devices are directed, the economy of the devices is such that there is no motivation to increase the complexity of the device or its operation, for the sake of allowing the device to be use continuously more than one crop / harvest cycle. * "In addition, the sterile conditions outside the Removable bioreactor devices are not needed or are not possible in many cases and thus once opened to extract the collected production is not cost effective, practical or possible, keep sterile opening, leading to contamination of the bag and any content can 'I stay inside. In this way, these disposable devices have no additional use after a cycle of I. 11 culture. Thus, removable bioreactor devices are relatively inexpensive for the quantities and volumes of production typically required by non-industrial users and are relatively easy to use by non-professional personnel. In fact, it is this aspect of simplicity of use and economic costs! I lower, which refers to the lower production volumes of removable devices, is' -ie, a greater attraction of removable bioreactor devices. In this way, removable bioreactor devices of the prior art have very little in common - i with industrial-scale bioreactors - operational structural or economies of scale - and in fact teach far away from providing a solution to the problems associated with .'the bioreactors on an industrial scale, rather any form of description or suggestion to such a solution. Therefore, the present invention represents a revolutionary solution to the above-mentioned problems, providing a disposable bioreactor device for the large-scale production of cell / tissue cultures. The device of the present invention, although eliminable, is characterized in that it comprises a reusable collection outlet orifice to allow the collection of at least a portion of the medium containing cells and / or tissues when desired, thus allowing the device to be collected. be used continuously for one or more consecutive, subsequent crop / harvest cycles. In an industrial environment, the sterility of the collection outlet orifice during and after harvesting can be ensured to a significantly high degree at relatively low costs by, for example, providing a sterile cover, in which all connections and disconnections necessary services. and from the device can be made.

When eventually the device is not contaminated, then it can be eliminated. Such devices can be manufactured economically, even for the production of volumes of 50 liters or more of culture. In addition, the ability to execute a number of crop / harvest cycles is economically profitable, further decreasing the cost per device. A group of such devices can be installed economically and the number of devices in the group can also be controlled to closely match production with demand. In this way, the transition from bioreactors from the pilot plant to large-scale production could also be achieved in a relatively inexpensive and simple way, by adding more devices to the group. In addition, the relatively low production volume of each device, coupled with the lack of solid mixers, results in relatively higher output compared to typical stainless steel bioreactors.

OBJECTS OF THE INVENTION It is an object of the present invention to provide an associated device and method for axonally culturing and harvesting cells and / or tissues and which does not have the aforementioned disadvantages. It is another object of the present invention to provide such a device, which is economical to produce and simple to use. Another object of the present invention is to provide such a device, which is removable, but nevertheless can be used continuously for a plurality of consecutive cycles of culture and collection of desired cells and / or tissues. Another object of the present invention is to provide such a device, wherein only the inoculant is required to be provided for the first cultivation cycle, while the inoculant for the subsequent cycles is provided by a portion of the culture broth, which remains in the device after collection thereof in a cycle preceding. Other . object of the present invention is to provide a group of such devices for the production on an industrial scale of cells and / or tissues. SUMMARY OF THE INVENTION A removable device and corresponding method for axenically growing and harvesting cells and / or tissues in at least one cycle, said device comprising a removable translucent and / or transparent sterilizable container having an upper end and a lower end, which The container can be filled at least partially with a suitable sterile biological and / or tissue and / or axenic inoculant and / or sterile air and / or other required sterile additive cell culture medium and / or tissue, said container comprising: a gas outlet means for removing excess air and / or waste gases from said container; an additive input means for introducing said inoculant and / or said culture medium and / or said additives into said container and characterized in that it further comprises a reusable collection means comprising an appropriate flow control means, to enable the collecting at least a desired portion of said medium containing cells and / or tissues when desired, thereby allowing said device to be used i continuously for at least one additional sequential crop / harvest cycle, wherein a remainder of said Medium that contains the remaining cells and / or tissues of a previously harvested cycle, can serve as an inoculant for a next crop and harvest cycle, where! said culture medium and / or said required additives are provided. Said device may further comprise an air-entraining means for introducing sterile air in the form of bubbles in said culture medium, through an inlet opening. The medium and air and other required additives are provided in suitable amounts during each cycle to allow the cultivation of said cells and / or losses from said inoculant. Said device can be eliminated when it is contaminated. In a second aspect of the invention, a group of these devices, suitably interconnected, allows the scale of cell / tissue production to be adjusted as required. J | DESCRIPTION OF THE FIGURES Figures la and lb illustrate the main components of a preferred embodiment of the present invention in front elevation view and in transverse side view, respectively. Figures 2a and 2b illustrate the main components of a second embodiment of the present invention in front elevation view and in transverse side view, respectively. Figure 3 illustrates the main components of a third embodiment of the present invention in transverse side view. Figure 4 illustrates the joints of the preferred embodiment of the present invention in front elevation. - Figure 5 illustrates the main components of a preferred embodiment of the group of the present invention. DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a disposable device for axenically culturing and harvesting cells and / or tissues in at least one cycle, said device comprising a removable translucent and / or transparent sterilizable container having an upper end and an end bottom, whose container can be filled at least partially with a culture medium of suitable sterile biological cells and / or tissues and / or axenic inoculant and / or sterile air and / or other sterile additives,? required, said container comprising: (i) a gas outlet means for removing excess air and / or waste gases from said container; (ii) j (an additive input means for introducing said inoculant and / or said culture medium and / or said additives into said container, and characterized in that it further comprises (iii) a reusable collection means comprising a control means suitable flow rate, to * 'allow the collection of at least a desired portion of said medium containing cells and / or tissues when desired, thereby allowing said device to be used continuously by at least one consecutive crop / harvest cycle, additional, where a remnant of said medium containing cells and / or tissues remaining from a previous harvested cycle, can serve as an inoculant for a next crop and harvest cycle, wherein said culture medium is provided and / or said required additives The present invention further relates to such a device further comprising an air inlet means for introducing sterile air in the form of bubbles in said culture medium through a first inlet opening, said air inlet means being connected to a suitable air supply. In this way, with reference to Figures 1, 2 and 3, corresponding respectively to preferred embodiments of the device, second and third, the device generally designated as (10), comprises a transparent and / or translucent container (20), having one end upper (26) and a lower end (28). Said container (20) comprises a side wall (22), which preferably is substantially cylindrical, although other shapes such as rectangular or polyhedral, for example, may also be suitable. Preferably, said lower end (28) is suitably formed to minimize sedimentation therein. For example, in the preferred embodiment, said lower end (28) is substantially frustoconical or at least comprises inclined walls ha? I-a above. In the second embodiment, the lower end (28) comprises a wall inclined towards f above (29). In the third mode, the lower end (28) is substantially cylindrical or alternatively convex. The aforementioned configurations of the lower end (28), together with the location of the outlet orifice (76) (described below) near the lower end (28), allows air to be supplied through said outlet orifice (76). ) to induce a mixing movement to the contents of the container at the lower end (28), which effectively minimizes sedimentation therein. However, the lower end may be substantially planar in other embodiments of the present invention. The container (20) comprises an internal fillable volume (30), which is typically between 5 and 50 liters, although said device (10) can also alternatively have an internal volume greater than 50 liters or less than 5 liters. 1 said internal volume (30) can be filled with a suitable sterile biological, cell and / or tissue culture medium (65) and / or axenic inoculant (60) and / or sterile air and / or other required sterile additives such as , antibiotics or fungicides, for example, as described below. In the aforementioned embodiments, the container 20 is substantially non-rigid, preferably being made of a non-rigid plastic material selected from the group comprising polyethylene, polycarbonate, a polyethylene copolymer and nylon PVC and EVA, for example. Optionally, the container (20) can be made from a laminate of more than one layer of said materials. As T is shown for the third embodiment in Figure 3, said container (20) may optionally comprise two concentric outer walls (24) to increase the mechanical strength and to minimize the risk of contamination of the contents, through the walls - - ^ of the container. In preferred embodiments, second and third, said device (10) is for aerobic use. In this way, the container (20) further comprises an air inlet means for introducing sterile air in the form of bubbles (70) in said culture medium (65), through an air inlet opening (72). . In the above-mentioned embodiments, said air inlet means comprises a pipe (74) which is connected to a suitable air supply (not shown) and which extends from said inlet opening (72) to a location within said container (20) at a distance di from the lower part of said lower end (28), where di may typically be about 1 cm, although it may be greater or less than 1 cm. Said conduit (74) can be made of silicon or other suitable plastic material and is preferably flexible. In this way, the duct (74) comprises an air outlet orifice (76) of suitable diameter to produce air bubbles (70) of a required average diameter. These bubbles not only aerate the medium (65), but also serve to mix the contents t of the container, thus also minimizing the settling at the lower end (28), as described above. The size of the bubbles supplied by the air inlet means will vary according to the use of the device, varying reasonably from below 1 mm to above 10 mm in diameter. In some cases, particularly in relation to plant cells, small bubbles can actually damage the cell walls and a bubble with a mean diameter of not less than 4 mm substantially outweighs this potential problem. In other cases, much smaller bubbles are beneficial and a sprinkler can be used in the air outlet hole (76) to reduce the size of the bubbles. In still other cases, air bubbles with a diameter of 10 mm or even larger can be optimal. Optionally, said outlet orifice (76) may be limited in position at said lower end (28) by means of a latch (not shown) or other means known in the art. In other embodiments, said device (10) is for aerobic use and thus does not comprise said air inlet means. Said container (20) further comprises a gas outlet means for removing excess air and / or residual gases from said container (20). These gases are collected in said upper end (26) of said container (20). Said gas outlet means may comprise a conduit (90) having a gas orifice., inlet (96) at or near said upper end (26), at a distance d4 from the bottom of said lower end (28), wherein d4 is typically 90 cm for the preferred embodiment. Said conduit (90) can be made of silicon or other suitable plastic material and is preferably flexible. Said conduit (90) is connected to a suitable exhaust means (not shown) by known means. Said exhaust means further comprises means, such as a filter or single pass valve suitable, for example, to substantially prevent the introduction of contaminants into said container through said gas outlet means. At least a portion of the upper end (26) can be suitably configured to facilitate the collection of waste gases before being withdrawn through said inlet orifice (96). Thus, in the preferred embodiments, first and second, the upper portion of the upper end (26) tapers progressively to a minimum cross-sectional area near the location of said inlet orifice (96). Alternatively, at least the upper portion of the upper end (26) may be correspondingly, substantially frusto-conical or convex. Said container (20) further comprises an additive input means for introducing said inoculant and / or said culture medium and / or said additives into said container. In the aforementioned embodiments, said additive input means comprises a suitable conduit (80) having | an outlet hole (86) preferably at or near said upper end (26), at a distance d3 from the bottom of said lower end (28), wherein d3 for the preferred embodiment is typically approximately 68 cm. Said conduit (80) can be made of silicon or other suitable plastic material and is preferably flexible. Said conduit (80) is connected by known means to a suitable sterilized supply of said inoculant and / or said culture medium and / or said additives. Said additive input means further comprises means for substantially preventing the introduction of contaminants into said container, through said additive input means and comprises, in these embodiments, a suitable single-pass filter or valve (84). Typically, the content level of the container (20) remains below the level of said exit orifice (86). Said container (20) further comprises a reusable collection means for collecting at least a first desired portion of said medium containing cells and / or tissues when desired, thereby allowing said device to be used continuously by at least one cycle of Subsequent cultivation. A second remaining portion of said medium containing cells and / or tissues serves as an inoculant for a next crop and harvest cycle, wherein said culture medium and / or said required additives are provided. Said collecting means can also be used to introduce the original volume of inoculant into the container, as well as to allow the collected material to flow therethrough and out of the container. In the above-mentioned embodiments l1, said collection means comprises a conduit (50) having an inlet (52) in communication with said internal volume (30) and an outlet orifice (56) outside said container (20) . Said conduit (50) can be made of silicon or other suitable plastic material and is preferably flexible. Said conduit (50) is of a relatively large diameter, typically about 2 cm, since the flow of cells and / or tissues collected therethrough may contain aggregates of cellular particles that can clog the narrowest passages. Preferably, said inlet (52) is located near the lower end (28) of said container (20), so that only the contents of the container above said inlet (52) are collected. In this way, at the end of each collection cycle, said second portion of the medium containing cells and / or tissues automatically remains at said lower end (28) of said container (20), up to a level below the level (51) of said orifice of entrance (52), which is at a distance d2 from the lower part of said lower end (28). Typically, d2 is approximately 25 cm for the preferred embodiment. Alternatively, said inlet orifice (52) may be located at the lowermost point in said container (20), where the operator could manually ensure that a suitable portion of the cell-containing medium and / or tissues remains in the container (20_). ) after collecting a desired portion of the medium and cells and / or tissues. Said collecting means further comprises a flow control means such as a suitable valve (54) and / or an aseptic connector (55) to block and to allow the flow of material ~ towards or out of said container (20) through said means of collection. Typically, said aseptic connector (55) is made of stainless steel and many examples thereof are known in the art. Preferably, said collecting means further comprises a means of prevention against contaminants to substantially prevent the introduction of contaminants into said container, through said collection means after collection. In the preferred embodiments, second and third, said pollution prevention means comprises a fluid separator (300). Said fluid separator (300) is preferably in the form of a substantially hollow U-shaped tube, an arm of which is mounted in the outlet orifice (56) of said collection means and the other arm having an external opening (58). The harvested cells / tissues can flow out of the device (10) through said collecting means, said fluid separator (300) and said opening (58), to then be collected in a suitable receiving container U, as described later. After the collection is completed, the air could possibly be introduced into the collection medium through the opening (56), accompanied by some reflux of collected material, thereby potentially introducing contaminants into the device. Said U-tube (300) substantially overcomes this potential problem by separating some of the collected material, i.e., cells / tissues downstream of the opening (56) thus preventing air and possible contaminants from entering the collection medium. Once the collection means is closed through said valve (54), the U-tube (300) is removed and typically sterilized for the next use or discarded. (Said U-tube (300) can be made of stainless steel or other suitable rigid plastic materials In the above-mentioned embodiments, said second remaining portion of said medium containing cells and / or tissues, typically comprises between 10% and 20% of the original volume of said culture medium and said inoculant, although said second portion may be greater than 20%, up to 45% or more, or less than 10%, below 2.5% or less, of said original volume, if Said device (10) optionally further comprises a joining means for joining the same to a cantilevered support structure I. In the above-mentioned embodiments, said support structure may comprise a bar (100) (Figures 1, 2) or rings (not shown) In the third embodiment, said attachment means may comprise a hook (25), preferably integrally attached to said upper end (26) of said container (twenty) . Alternatively and as shown for the preferred and second embodiments, in Figures 1 and 2, respectively, said attachment means may comprise a preferably flexible and substantially cylindrical clamp (27) of suitable material, typically of the same material as used for the container (20) either integral with or substantially joined (through fusion welding, for example) to the upper end (26) of the device. Said container (20) can be formed by fusing together two suitable sheets of suitable material, as exemplified above, along predetermined joints. Referring to Figure 4, the two sheets (200) of material can be cut into an approximately elongated rectangular shape and juxtaposed one on top of the other. The sheets are then joined together by melting in a manner well known in the art, to form the joints along peripheries (205) and (206) of the two longer sides and along the periphery of one of the ends shorter (210) and again parallel and internally displaced thereto to form a gasket (220) at the upper end of the container (20). The fusion welded joints (207) and (208) along the long sides and placed between these parallel short end joints (210) and (220), they can be cut or otherwise removed, effectively leaving a clamp of material (27). The lower end (28) of the container (20) is formed by fusing the remaining short end of the sheets along two lines of inclined joints (230) and (240), which mutually converge from the joints ( 205) and (206) ll of the long sides. Optionally, the two lines of inclined joints (230) and (240) can be joined above the vertex by other lines of fusion-welded joint (260) approximately orthogonal to the long side joints (205) and (206). Before fusion welding of the two sheets together, the rigid plastic nails (270), (290), (280) and (250) can be melt melted at locations corresponding to said air inlet means, said gas outlet means, said additive input means and I 'said collection means, respectively. These nails provide mechanical attachment points, suitable for each of the corresponding entry and exit means. In all embodiments, the device (10) is made of a material or materials that are biologically compatible and that allow the container to be sterilized before being used first. The present invention also relates to a group of disposable devices for axenically culturing and harvesting cells and / or tissues in cycles, wherein each of a plurality of these devices is structurally and operationally similar to said device i (10), defined and previously described. Referring to Figure 5, a group (500) • i comprises a plurality of said devices (10), which are held in a structure or structures (not shown) by means of said joining means. Typically, the group can be divided into a number of groups, each group comprising a number of devices (10). In the preferred embodiment, said air inlet means of the devices (10) are interconnected in each group. In this way, said air inlet ducts (74) of each device (10) of the group are connected to a common pipe (174) having a free end (170), which is provided with an aseptic connector (175) . The sterilized air is provided by a suitable air compressor '(100) having a suitable sterilization means (110) such as one or more filters. The compressor (100) comprises a supply conduit (101) having an aseptic connector (176) at its free end, which is typically connected to said aseptic connector (175) located at the free end of the common pipe (174) . This . Connection is made at the start of each series of growth / harvest cycles in a mobile sterile cover (380) to ensure that sterile conditions are maintained during the connection. The sterile cover (380) provides a relatively simple lower cost system for connecting the various services such as, air, media, inoculant and harvested cells, to and from the group i of devices (10) under substantially sterile conditions. . Similarly, at the end of each series of growth / harvest cycles, the connectors (175) and '(176) are disconnected in the sterile cover (380) and the used devices are discarded, allowing the connector (175) at the end of the compressor to connect to the connector (176) of a new group of devices. The sterilized air is typically provided continuously or alternatively in predetermined pulses, during each culture cycle. In the preferred embodiment, excess air and / or waste gases from each of said devices (10) is withdrawn into the atmosphere through the common pipe (290) suitably connected to each gas outlet means. corresponding (90). Said common pipe (290) is provided with suitable means (210), such as one or more filters, to prevent contaminants from flowing into said devices (10). Alternatively, the gas outlet means (90) of each device (10) can be individually allowed to vent to the atmosphere, preferably through suitable filters that substantially prevent contaminants from flowing into the device (10). The media and additives are contained in one or more retainer tanks (340). For example, the micro elements, macro elements and vitamins can be kept in different deposits, while the additives such as, antibiotics and fungicides can also be maintained in .1 still other separate deposits. The pump means (345) serving each tank allows the desired relative proportions of each component of the media and / or additives to be supplied at a controllable and predetermined flow rate to a static mixer (350), or through the which water, whether distilled or purified and suitably filtered, flows from an adequate supply (360), preferably with the aid of a suitable pump means (365) (Figure 5). By adjusting the flow rates of the pumping means (345) and (365), for example, the concentration of the media as well as the additives available to be supplied to said device (10) can be controlled. The media and / or additives mixed with water can then be supplied from said static mixer (350) under sterile conditions through a filter (310) and a supply conduit (370) having an aseptic connector (375), as its end free (390). In the preferred embodiment, the input of the additive conduit (80) of each corresponding device 1 (10) in the group of said devices is interconnected through the common pipe (180), which comprises at its free end an aseptic connector common (376). Said common aseptic connector (376) can then be connected, in said sterile cover (380), to the aseptic connector (375) at the free end (390) of the additive means and conduit (370), thus allowing each device (10) ) of the group is supplied with means and additives. At the end of the life of the devices (10) and before discarding them, the aseptic connectors (375) and (376) are disconnected in the sterile cover J. The aseptic connector (375) is then ready to be connected to the new aseptic connector (376) of the next sterilized group of new devices (10) of the group, ready for the next series of crop / harvest cycles. The sterile cover (380) can also be used to connect the media / additive reservoir (350) to each of a number of device groups (10) in the group, in turn, during the lifetimes of the devices in these groups. Thus, when a group of devices has been served with media / additives, the aseptic connector (376) of this group is aseptically sealed temporarily in the sterile cover (380), which then moves to the next group. of devices where their common aseptic connectors (376) are connected to the sterile connector (375) of the conduit (370), thus allowing this group of devices to be served with means / additives. In another embodiment, said movable sterile cover (380) can be used to connect together the free end (390) of a preferably flexible supply line .1 connected to said static mixing tank (350), in turn, to the input means. of additives of each device (10). Said sterile cover (380) can then be moved from said device (10) to the next one, each time said end (390) is connected to the input end of the corresponding conduit (80), in order to allow the means to be provided at the same time. every device.

The sterile cover (380), together with the aseptic connecting means, preferably made of stainless steel, at said end - (390) and the inlet opening of the conduit (80) of the corresponding device (10), respectively, allow each device (10) is connected and disconnected subsequently to the end (390) and in this way to the supply of means, under sterile conditions. Many other examples of suitable connecting means for connecting two conduits together are well known in the art. The. Suitable filters are provided at the end (390) and in the conduit (80), respectively, to .1 avoid or at least minimize potential contamination of the contents of the container. The sterile cover (380) can thus automatically or manually move the device (10) to the device (10) and in turn on each device, an operator can connect the device (10) to the media supply using the cover sterile • I (380), fill the device with an adequate amount of i 'media and / or additives and subsequently disconnect the sterile cover (380) from the device, then: l move it to the next device. Of course, the extreme (390) can be adapted to comprise a plurality of connecting means (375) instead of only a single sterilized connecting means (375), so that instead of one, a plurality of similar devices (10) having means The corresponding connection cables (376) can be connected simultaneously to the supply of means, through the trolley (3-80). Each time, before connecting said end (390) to i each device or group or group of devices, the corresponding connection means (375) and (376) are typically sterilized in autoclave. In yet another embodiment of the group, a single conduit or a set of conduits (not shown) connects said static mixer (350) to said device (10) or to a corresponding set of devices (10), respectively, at a time, wherein a conveyor system transports the device (10) or set of devices (10) to said single conduit or set of conduits, respectively or vice versa. After filling said device (10) or set of devices (10), the conveyor allows an additional device (10) or set of devices (10) to be connected to the static mixer (350) by means of said single conduit or assembly. conduits, respectively. In the preferred embodiment, the means of collecting; each of the devices (10) of each group are interconnected. In this way, the collection conduits (50) of each of said devices (10) are connected to a common collection pipe (154) having a free end (150), which is provided with an aseptic connector (155). ). Preferably, each of said collection conduits (50) may comprise a valve (54), as described above, to block or allow the flow of cells collected from each corresponding device (10). In this way, for example, if it is determined that a number of devices in a particular group is contaminated, while the other devices do not, then the cells of these latter devices can be collected without fear of contamination of the previous devices, provided that valves' _ (54) of contaminated devices remain closed. Preferably, said common pipe further comprises a common shut-off valve (259) upstream of said aseptic connector (155). Preferably, said pollution prevention means is provided to substantially prevent the introduction of contaminants into said container, through said collection means after collection. In the preferred embodiment, said pollution prevention means __ comprises a substantially U-shaped fluid separator (400), having an aseptic connector (156) on one arm thereof, the other arm having an opening (158) in fluid communication with a reception tank (590). The aseptic connectors If (155) and (156) are then interconnected in said movable sterile cover (380) under sterile conditions. The collection is then carried out by opening the valves (54) of all the devices in the group that are not contaminated, as well as the common valve (259). The cells of the groups will then flow into the reception tank (590), preferably under gravity, although in some cases a suitable pump may be used. After the collection is completed, the aseptic connectors (155) and (156) can be disconnected in said sterile cover (1) (380), which can then be moved to the next group of devices (10): the corresponding aseptic connector ( 155) of this group can then be interconnected with the aseptic connector (156) of the U-conduit (400) and thus allows the cells of this group of devices to be collected. In another embodiment, a single conduit or a set of conduits (not shown) can connect said common receiving reservoir to said device (10) or to a corresponding set of devices (10), 1 respectively at a time, wherein a system The conveyor transports the device (10) or set of devices (10) to said single conduit or set of conduits, respectively or vice versa. After the collection of said device (10) or set of devices (10), the conveyor allows an additional device (10) or - set of devices (10) to be connected to said common reception tank, by means of said single conduit or .1 set of conduits, respectively. In another embodiment, each device (10) can be collected individually, wherein said collection means of each device comprises said pollution prevention means to substantially prevent the introduction of contaminants into said container, through said collection means after The recollection. In this embodiment, said pollution prevention means comprises said U-shaped fluid separator (400), as described; !! above, having an aseptic connector (156) on one arm thereof, the other arm having an opening (158) in fluid communication with a reception tank (590).

Said means of collection comprises an aseptic connector (55), which can be connected to the aseptic connector (156) of the fluid separator (400) in said movable sterile cover (380) under sterile conditions. The collection is then effected upon opening the valve (54) of the device, where the cells will then flow into the receiving reservoir, preferably under gravity, *? although in some cases an appropriate pump can be used. After the collection is completed, these aseptic connectors (55) and (156) can be disconnected in said sterile cover (380), which can then be moved to the next device (10): the corresponding aseptic connector (55) of the medium of collection of this device, then it can be interconnected with the aseptic connector (156) 'of the U-duct (400) and thus allows the cells of this next device to be collected. In the preferred embodiment, said collection means can also be used to initially provide the inoculant at the start of a new series of growth / harvest cycles. In this manner, the in-mixing with the sterilized medium in a suitable reservoir having a supply conduit comprising at its free end an aseptic connector that connects to said aseptic connector (155) of the common collection line (154) in said sterile cover (380). The inoculant can then be allowed to flow under gravity or with the aid of a suitable pump, to each of the devices (10) of the group, through said common collection pipe (154), after which the aseptic connectors. they are disconnected in the sterile cover. Alternatively, said inoculant can be introduced into the devices through said additive input means, in particular said common line of the additive medium (180), in a manner similar to that described above, considering the collection means and the delivery conduit. common collection (155), mutatis mutandis. The present invention also relates to a method for cultivating and collecting cells and / or tissues in a disposable device of multiple uses comprising the steps of: a)? (Roporcionar said device (10), defined above; b) providing sterile air to said container, through said air inlet means i during each cycle either continuously or in pulses; c) providing said sterile culture medium and / or said sterile additives, through said additive input means; d) providing a non-toxic inoculant, through said collection means; 10 e) optionally illuminate said container with • I external light means; f) allowing said cells and / or tissues j to grow in said medium to a desired production, g) continuously allow excess air and / or residual gases to leave said container, through said outlet means of gases; h) verify the contaminants and / or the quality of the cells / tissues that are produced in said container: if contaminants are found The present or the cells / tissues that are produced are of poor quality, the device and its contents are eliminated; if no contaminants are found, stage i) is executed; i) collecting at least said first portion Desired from said medium containing cells and / or tissues, while leaving said second remaining portion of the medium containing ri and / or tissue cells in said container, wherein said second portion of the medium can serve as an inoculant for a subsequent crop / harvest cycle. j) providing said sterile culture medium and / or said sterile additives for the following .1 crop / harvest cycle, through said additive input means; *. k) repeating steps b), e), f), g), h), i) and j) i I a plurality of times until in h) i? 'I find such contaminants present or the cells / tissues that are produced are of poor quality, after which the device and its contents are removed. The present invention also relates to a method for culturing and axenically harvesting cells and / or tissues in an ancrobical manner in a group of removable devices, comprising the steps of: a) providing a group (500) of at least one group of said devices (10), wherein said devices do not comprise air inlet means and by said at least one device (10) thereof; b) providing axenic inoculant to said device, through said common collection line; c) providing said sterile culture medium and / or said sterile additives to said device, through said inlet pipe. f of common additives; d) optionally illuminating said device with external light means; 10 e) allowing said cells and / or tissues in said device to grow in said medium to a desired production; ti f) allow excess air and / or residual gases to leave said device continuously, through said common gas outlet pipe; g) verify the contaminants and / or the quality of the "cells / tissues that are produced in said device: if contaminants are found in said device or the cells / tissues that are produced are of poor quality, said means of collection of said device is blocked, avoiding the contamination of other said *! devices of said group; yes in all of Said devices of said group are contaminants or the cells / tissues that are produced therein are of poor quality, all the devices and their contents are eliminated; if no contaminants are found and the quality of the cells / tissues produced is acceptable, the device is considered collectible and step h) is executed; h) for each said collecting device of step g), collecting at least said first The desired portion of said medium containing cells and / or tissues, through said common collection line and said contamination prevention means for a proper reception deposit, while leaving said The second portion of the medium containing cells and / or tissues in said container, wherein said second portion of the medium serves as an inoculant for a next crop / harvest cycle; I) providing said sterile culture medium and / or said sterile additives for the next crop / harvest cycle through said additive input medium; j) repeat steps d), e), f), g), h) and i) a A plurality of times until in g) said contaminants are found or the cells / tissues that are produced are of poor quality for all of said devices of said group, after which said pollution prevention means are disconnected from said means of • 1 common collection and such devices and their contents are eliminated. The present invention also relates to a method for culturing and axially harvesting cells and / or tissues aerobically in a group of removable devices, comprising the steps of: a) providing a group (500) of at least one group of said devices (10), wherein said devices comprise air inlet means as described hereinafter and by at least one of said devices (10) of. the same ones; \ > i b) providing axenic inoculant to said device, through said common collection line; c) providing said sterile culture medium ii and / or said sterile additives to said device, through said common additive inlet pipe; | d) providing sterile air to said device, through said common air inlet pipe; e) optionally illuminating said device with external light means; I f) allowing said cells and / or tissues in said device to grow in said medium at a desired production; g) allow excess air and / or residual gases • I to leave said device continuously, through said common gas outlet pipe; h) verify the contaminants and / or the quality of the cells / tissues that are produced in said container: if contaminants are found in said device or the cells / tissues that are produced are of poor quality, said means of,? • collection of said device is blocked, avoiding the contamination of other said devices of said group; yes in all of Said devices of said group are contaminants or the cells / tissues that are produced therein are of poor quality, all the devices and their contents are eliminated; If no contaminants are found and the quality of the cells / tissues produced is acceptable, the device is considered collectible and step i) is executed; * i) for each said collecting device of step h), collecting at least said first desired portion of said medium containing cells and / or tissues, through said common collection line and said means of prevention against contamination for a adequate reception deposit, while leaving said The second portion of the medium containing cells and / or tissues in said container, wherein said second portion of the medium serves as an inoculant for a next crop / harvest cycle; J) providing said sterile culture medium and / or said sterile additives for the next crop / harvest cycle, through said additive input means; ii) repeating steps d), e), f), g), h), i) and j) a plurality of times until in h) said contaminants are found or the cells / tissues that are produced are of poor quality for all of said devices of said group, after which said means of The prevention of contamination is disconnected from said common collection means and said devices and their contents are eliminated. Typically, a water purification system supplies pyrogen-free and deionized water to a reservoir comprising concentrated media and the diluted medium is then pumped to said device (10), via said additive inlet medium. The filters, typically 0.2 μm, are installed in the supply ducts and also just upstream of said additive input means to minimize the risk of contamination of the contents of the container in each device (10). Alternatively or additionally, a single-pass valve can also be used to minimize this risk. For • > the first cycle of culture of each device (10), the inoculant, typically a sample of the cell type that is required is collected in said device (10), pre-mixed with the media or water in a container sterilized by steam and it is introduced into the device (10), through the collection means. The means are then introduced into the device (10) through the said additive input means. For subsequent cycles, only the media and / or additives are introduced, such as? Described above,? 11 Typically, an air compressor provides substantially sterilized air to each said device? < (10), through a number of filters, a coarse filter to remove the particles, a moisture filter and dryer to remove moisture and a thin filter, typically 1 0.2 μm, to remove contaminants. Preferably, another filter just upstream of said air inlet means, further minimizes the risk of contamination of the contents of the container. For each said device (10), all connections to the container (20), that is, to said air inlet means, to said additive input means and preferably also to said gas outlet means and to the collection means , they are sterilized in an autoclave before being used, and sterility is maintained during connection to the peripheral equipment including, for example, said air supply and said exhaust means when executing the connections in the sterile cover, as described above. The temperature control for each device (10) is preferably provided by suitable air conditioning means. Optional illumination of the device may be provided by suitable fluorescent light means installed around said device (10), when required for cell-growth.

During each growing cycle of each device (10), - the contents of each corresponding container (20) are typically aerated and mixed by approximately 7 to approximately 14 days or more, under suitable lighting and temperature conditions. At the end of the culture cycle for each device '(10), said corresponding collection means is typically connected to a pre-sterilized environment by means of suitable connectors which are sterilized before and during connection, as described i < previously. Harvesting then takes place, leaving behind between about 2.5% to about 45%, although typically between about 10% to about 20% of the cells and / or tissues serve as an inoculant for the next cycle. The collected cells / tissues can then be dried or extracted, as required. The present invention will be described in more detail with reference to the following example, which is not intended to limit the scope of the invention. EXAMPLE Vinca T cell Culture A group of 10 bioreactors (each a device according to the invention) was used, each with an elaborate container of nylon-polyethylene copolymer, - (wall thickness 0.1 mm, 20 cm diameter, 1.2 m in height), complete with ports of 30 mm to 5 cm (for air intake means), 25 cm (for collection means), 68 cm (additive input means) and 90 cm (means of gas outlet) from the bottom; fillable volume approximately 10 liters. The bioreactors, together with their accessories, were sterilized by gamma irradiation (2.5 mRad). Í Nine liters of Schenk's mineral / vitamin medium & Hildebrandt, 2 mg / l each of chlorophenoxyacetic acid and 2,4-dichlorophenoxyacetic acid, 0.2 mg / l of cmetin, 3% of sucrose and 900 ml of packed volume inoculum of Catharanthus roseus (Vinca) cells line V24, introduced into each bioreactor. The volume of air above the surface of the medium was 3 1. The aeration was carried out using a flow volume of 1.5 l / min of sterile air, provided through a 4 mm orifice (air inlet means) ), located 1 cm from the bottom of the container. The bioreactors were mounted at a controlled ambient temperature (25 ° C) and the culture was continued for 10 days, until the packed volume was increased to approximately 7.51 (75% of the total volume, a double ratio of 2 days during the logarithmic phase). ). At this point, the cells were harvested by extracting 9 liters of medium and cells through the collection medium and 9 liters of fresh sterile medium were added along with the same additives, through the additive input medium. Cells were harvested again for intervals above 10 days, for 6 additional cycles, at which time the series was completed. In this way, a total weight of 6.5 cells! .1 > Recent (0.5, kg dry weight) were collected for seven periods of 10 days, from each of the 101 capacity bioreactors. These cells had a 0.6% total alkaloid content, the same as the start line. Although only some embodiments have been described in detail in the foregoing description, the present invention is not limited thereto and is only defined by the scope of the claims.

Claims (5)

1. NOVELTY OF THE INVENTION Having described the present invention, it is considered as a novelty and therefore the property described in the following claims is claimed as property. 1. A removable device to cultivate and I! collecting axemcally cells and / or tissues in at least one cycle, said device comprising a translucent and / or transparent sterilizable removable container having an upper end and a lower end, which container can be filled at least partially with a cell culture medium and / or biological, sterile, suitable and / or axenic inoculant and / or sterile air and / or other required sterile additives, said "container: (i) a gas outlet means for removing excess air, and / or waste gases from said container; (ii) an additive input means for introducing said inoculant and / or (said culture medium and / or said additives into said container, and characterized in that it further comprises (iii) a reusable collection means comprising a means of suitable flow control, to allow the collection of at least a desired portion of said medium containing cells and / or tissues when desired, thereby allowing said device to be used continuously by at least one additional consecutive crop / harvest cycle , wherein a remnant of said medium containing cells and / or tissues remaining from a previous harvested cycle, can serve as an inoculant for a next crop and harvest cycle, wherein said culture medium and / or said additives are provided.
2. 2. The device according to claim 1, characterized in that it further comprises an air inlet I for introducing sterile air in the form of 1 burb. ujas in said culture medium, through a first entrance opening, wherein said input means of i! air is connected to an adequate air supply.
3. 3. The device according to claim 1 or claim 2, characterized in that said collection means comprises prevention means against contamination to substantially prevent the introduction of contaminants in said container, through said collection means.
4. 4. The device according to any of the preceding claims, characterized in that said container is not rigid.
5. 5. The device according to any of the preceding claims, characterized in that said container is made of a non-rigid plastic material. The device according to claim 5, characterized in that said material is selected from the group comprising polyethylene, polycarbonate, a copolymer of polyethylene and nylon, PVC and EVA. 7. "The device according to claim 5 or 6, characterized in that said container is made of a laminate of more than one layer of said materials." 8. The device according to any of claims 5 to 7, characterized in that said container is formed by melt-bonding two suitable sheets of said material along predetermined joints 9. The device according to any of the preceding claims, characterized in that said air inlet means comprises an air inlet duct extending therefrom. opening of entry to a location within said container at or near said lower end thereof 10. The device according to any of the preceding claims, characterized in that at least some of said air bubbles comprise an average diameter of between about 1 mm. and approximately 10: mm mm 11. The device according to any of the preceding claims, c characterized in that at least some of said air bubbles comprise an average diameter of about 4 mm. 1 - . 12. The device according to any of the preceding claims, characterized in that said container comprises a suitable filter mounted on said gas outlet means to substantially prevent the introduction of contaminants into said container., through said gas outlet means. The device according to any of the preceding claims, characterized in that said container further comprises a suitable filter mounted on said additive input means to substantially prevent the introduction of contaminants into said container, through said additive input means. . 14. The device according to any of the preceding claims, characterized in that said means for preventing pollution comprises a U-shaped fluid separator, wherein an arm thereof is assembled aseptically to an external outlet orifice of said medium. collection by means of suitable aseptic connection means. 15. Device according to any of the preceding claims, characterized in that said f1 collection means is located in the lower part of said lower end of said container. device according to any of claims 1 to 14, characterized in that said collection means is located near the lower part of said lower end of said container, such that at the end of each collection cycle said remnant of said means that contains cells and / or tissues automatically remains at said lower end of said container to a level below the level of said collection means, 17. * The device according to any of the preceding claims, characterized in that said remnant of said medium containing cells and / or fabrics comprises between about 2.5% and about 45% and preferably between about 10% and about 20% of the original volume of said culture medium and said inoculant. 18. The device according to any of the preceding claims, characterized in that said lower end is substantially convex. 19. The device according to any of claims 1 to 17, characterized in that said lower end is substantially frustoconical. 20. The device according to any of the preceding claims, characterized in that said container comprises an internal fillable volume of between approximately 5 liters and approximately 50 liters. The device according to any of the preceding claims, characterized in that said device further comprises suitable joining means for attaching the same to a suitable support structure. u 22. The device according to claim 21, characterized in that said connecting means comprises a clamp of material preferably integrally connected to said upper end of said container 23. A group of said devices according to any of the preceding claims. characterized in that it comprises at least two of said removable devices 24. The group according to claim 23, characterized * in that said devices are supported by a suitable support structure through said joining means of each said device. claims 23 or 24, characterized in that said gas outlet means of each said device is suitably connected to a common gas outlet pipe, which optionally comprises a means suitable to prevent contaminants from flowing to said devices. 26. The group according to claim 25, characterized in that said edio to prevent such pollutants from flowing to said devices comprises a suitable filter. 27. The group according to any of claims 23 to 26, characterized in that said input means of; Additives for each said device is connected in a suitable manner to a common additive inlet pipe having a free end optionally comprising suitable aseptic connection means therein. 28. The group according to claim 27, characterized in that said free end is connected to an adequate supply of means and / or additives 29. The group according to any of claims 23 to 28, characterized in that said collecting means of each said device is suitably connected to a common collection line having a free end optionally comprising a suitable aseptic connection means therein 30. The group according to claim 29, characterized in that it further comprises a means of prevention against contamination to substantially prevent the introduction of contaminants in said container, through said common collection pipe 31. The group according to claim 30, characterized in that said means of prevention against pollution includes a fluid separator in the form of U, wherein μn arm thereof is free having an opening and where the other end thereof is mounted aseptically to said free end of said common collection pipe, through a suitable aseptic connection means. 32. The group according to claim 31, characterized in that said free end of said U-pipe is connected to a suitable receiving tank. The group according to any of claims 23 to 32, characterized in that said air inlet means of each said device is suitably connected to a common air inlet pipe having a free end optionally comprising an aseptic connection means in the same. 34. The group according to claim 33, characterized in that said free end is connected to a suitable air supply. 35. A method for axenically culturing and harvesting cells and / or tissues in a removable device, comprising the steps of: a) providing said device comprising a removable translucent and / or transparent sterilizable container having an upper end and a lower end, which container can be partially filled with a biological, sterile, suitable and / or axenic inoculant and / or sterile air and / or other sterile additives required cell culture medium and / or tissue culture medium, said container comprising: (i) a gas outlet means for removing excess air and / or residual gases from said container; (ii) a means of introducing additives to introduce said inoculant and / or said culture medium and / or said additives into said container; (iii) a reusable collection means comprising a suitable flow control means, to allow the collection of at least a portion of said medium containing cells and / or tissues when desired, thereby allowing said device to be used continuously by at least one additional consecutive cycle, wherein a remnant of said medium containing cells and / or tissues remaining from a previously harvested cycle, can serve as an inoculant for a next crop and harvest cycle, wherein said medium is provided. cultivation and / or said additives required; b) providing axenic inoculant, through said collection means; c) providing said sterile culture medium and / or said sterile additives, via said additive input means; d) optionally illuminating said container with external light means; I, t e) allowing said cells and / or tissues to grow in said medium at a desired production; f) allowing the excess? i of air and / or residual gases to leave said container t continuously, "through said gas outlet means; g) verify the contaminants and / or the quality of the cells / tissues that are produced; in said container: if contaminants are found or the cells / tissues that are produced are of poor quality, the device and its contents are eliminated, if no contaminants are found, step h) is executed, h) collecting said desired portion of said medium containing cells and / or tissues, while leaving said remnant of the medium containing cells and / or tissues in said container, wherein said remnant of the medium serves as an inoculant for a next cycle of, 1 culture / harvest; said sterile culture medium and / or said sterile additives for the next crop / harvest cycle, through said additive input means; k) repeating steps d), e), f), g), h) and i) ) a ..plurality of vec it is until in g) that the contaminants are found or the cells / tissues that are produced are of poor quality, after which the device and its contents are eliminated. 36. The method according to claim 35, w characterized in that said device further comprises an air inlet means for introducing sterile air into the suitable sterile air, said method further comprises the step of providing sterile air to said air inlet means during the first and each subsequent cycle. S V - 37. tel method according to claim 36, characterized in that said sterile air is supplied continuously during at least one culture cycle, 38. The method according to claim 36, characterized in that said sterile air is supplied in pulses during at least one culture cycle. 39. A method for axenically culturing and harvesting cells and / or tissues in a group of removable devices, comprising the steps of: a) providing a group of devices according to claim 31 and at least one of said devices thereof; b) providing axenic inoculant to said device, through said common collection line; c) providing said sterile culture medium and / or said sterile additives to said device, through said common additive inlet line; d) illuminate! optionally said device with external light means; e) allowing said cells and / or tissues in said devices to grow in said medium at a desired production; f) allowing excess air and / or residual gases to discharge said device continuously, through said common gas outlet pipe; g) fl verify the contaminants and / or the quality of the I cells / tissues that are produced in said device: if contaminants are found in said device or the cells / tissues that are produced are of poor quality, said means of collection of said device is blocked, avoiding contamination of other said devices of said group; if in all of said devices of said group there are contaminants and / or the cells / tissues that are produced in them are of poor quality, all the devices and their contents are eliminated; if no contaminants are found and the quality of the cells / tissues produced is acceptable, the device is considered collectible and step h) is executed; h) for each said collecting device of step g), collecting said desired portion of said medium containing cells and / or tissues, through said common collection line and said means of prevention against contamination. suitable, while leaving said remnant of the medium containing cells and / or tissues in said container, wherein said second portion of the medium serves as an inoculant for a next crop / harvest cycle; i) providing said sterile culture medium and / or said sterile additives for the next crop / harvest cycle, through said additive input means; j) repeating steps d), e), f), g), h) and i) a plurality of times until in g) said contaminants are found or the cells / tissues that are produced are of poor quality for all of said devices of said group, after which said pollution prevention means are disconnected from said common collection means and said devices and their contents are eliminated. 40. A method for axially culturing and harvesting cells and / or tissues aerobically in a group of removable devices, comprising the steps of: a) providing a group of devices according to claim 34 and at least one of said devices thereof; b) providing axenic inoculant to said device, through said common collection line; c) providing said sterile culture medium and / or said sterile additives to said device, through said common additive inlet line; d) providing sterile air to said device, through said common air inlet pipe; e) optionally illuminating said device with external light means; f) allowing said cells and / or cells in said device to grow in said medium at a desired production; g) allowing excess air and / or residual gases to leave said device continuously, through said common gas outlet pipe; h) verifying the contaminants and / or the quality of the cells / tissues that are produced in said I 'device: if contaminants are found in said device H or the cells / tissues that are produced are of poor quality, said means of collection of said device is blocked, avoiding contamination of other said devices of said group; if in all of said devices of said group there are contaminants and / or the cells / tissues that are produced in them are of poor quality, all the devices and their contents are eliminated; if no contaminants are found and the quality of the cells / tissues produced is acceptable, the device is considered harvestable and step i) is executed; i) for each said collecting device of step h), collecting at least a desired portion of said medium containing cells and / or tissues, through said common collection pipe and said pollution prevention means for a deposit of suitable reception, while leaving said remnant of the medium containing cells and / or tissues in said container, wherein said remnant of the medium serves as an inoculant for a next crop / harvest cycle; j) providing said sterile culture medium and / or said sterile additives for the next crop / harvest cycle, through said additive input means; k) repeating steps d), e), f), g), h), i) and j) a plurality of I times until in h) said pollutants are found or I are produced of poor quality for slides of said group, after which said pollution prevention means are disconnected * from said common collection means and said devices and their contents are eliminated. 1 SUMMARY A disposable device and a method for axenically culturing and harvesting cells and / or tissues in consecutive cycles. The device consists of a translucent, translucent and / or transparent sterilizable container that can be at least partially filled with a culture medium of suitable, sterile, biological, and / or axenic cells and / or tissues and / or sterile air and / or tissue. other sterile additives required. The container has a means for removing excess air and / or residual gases therefrom and a means for introducing the inoculant, and / or the culture medium and / or additives therein. The device is characterized by having a reusable collection means to allow the collection of at least one portion of the medium containing cells and / or tissues when (as desired), thereby allowing the device to be collected. A previously collected cycle can serve as an inoculant for a next crop and harvest cycle, by providing the culture medium and / or additives. In this way, the device can be used continuously in consecutive cycles and can be eliminated when contaminated. In a second aspect of the invention, a group of these devices, suitably interconnected, allows the production scale of 1 cells / tissues to be adjusted when required.