DE3228047A1 - Method for growing cell tissue - Google Patents

Description

** patenta'n walte "**" * Π

DR.-ING. R. DÖRING - 5 - DIPL.-PHYS. DR. J. FRICKE

BRAUNSCHWEIG MUNICH

The A.P.V. Company Limited Manor Royal, Crawley, Sussex, England

"Method for Growing Cell Tissue"

The invention relates to a method for growing cell tissues.

In the manufacture of cellular tissues, the cells become traditional grown in roux cylinders or roller bottles. In order to obtain large quantities of tissue, large numbers of such bottles are required necessary. This method is burdened with a high work unit. Many different systems have been proposed to help Cell cultures that require the cells to be anchored, such as animal, plant or fungal cells single-layered to grow on an adhesive surface, to convert from laboratory conditions to large-scale production. Lots of cell cultures show an anchoring dependency on a corresponding surface as a requirement for growth. There the contact from cell to cell prevents further growth so a single-layer cell culture is created, which the available area covered. For high production speeds of biochemical substances, for example "of vaccines, e.g. for mumps, for rubella, measles, flu, Paragrippe, Marek's disease or infections of the respiratory system

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- 6 pathways of cell lines, fibroblast from chick embryos, virus cells

of ox, green monkey cells, human diploid lung or! Skin fibroblasts, a large surface area is required.

The production of many virual vaccines and biochemicals e.g. interferon, human growth hormones, somatastatin, alpha i thymasin-1 and human insulin depend on the I

rich cultivation of animal cells in the form of a Surface adhering monolayer. Similar products can be obtained from plant cells and from fungal cell tissue cultures will. ;

After the techniques of molecular biology have produced many new rows of cells, production levels are increased
aimed at and processing devices offered, which
have larger surfaces.

In particular, a process unit is available which has a
Has series of metal disks, which together with a
Shaft are mounted rotating. These discs are in one
Boiler mounted, the supply and discharge lines for
has liquid and gaseous median. In operation it will!
Vessel filled with a serum and held so that the j
Disks are in · horizontal planes so that the
Cells can settle on the plates. The kettle is tilted so that the panes are in a vertical plane
are located, with the boiler being partially emptied, by one
Maintain headroom for ventilation. The discs will

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then slowly rotated so that the cells are ready for further treatment can be continued. In another embodiment of the apparatus, the cells are in a row , grown and treated from pipes.

Devices of this type are expensive to manufacture and cannot easily be adapted to production requirements will. Precise control of the process is also not easily possible, since the culture is essentially batch-wise : takes place, where the temperature and the other variables are not; can be easily controlled from the outside.

It is an object of the invention to provide a method of the above in more detail

designated type so that the identified after-; parts and difficulties are avoided.

; j

To carry out the method, the invention provides that

; the cells are placed on plates of a plate heat exchanger

[ lets settle and allows a growth medium to pass in continuous circulation through the flow spaces defined between the plates.

Advantageously, a virus or some other additive is added to the circulating growth medium during circulation. leads.

'By inoculating the growth medium with a virus, a

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Infection in place of the cells can be achieved what j

leads to the production of vaccines. The virus can too can be used to transform the cell types. Alternatively, the culture can also be a trasposon or a Plasmid can be added. In certain circumstances, other additives may be required, e.g. antibiotics or an additive like sodium butyrate to stimulate interferon production.

Preferably, air or oxygen is dissolved in the circulating growth medium upon entry into the heat exchanger.

In particular, means are provided to remove atmospheric oxygen or carbon dioxide from the growth medium after it has left the Remove the heat exchanger.

In one embodiment, only alternating flow spaces are used for the cultivation, while the intermediate, lying flow spaces are used for the circulation of water or another medium at a controlled temperature will.

In one embodiment, a selected variation is the effective area on which the animal cell tissues are grown are obtained by passing vaccine and growth media through selected flow spaces of the heat exchanger diverts.

By separating the heat exchanger into two or more separate growth areas, the initial vaccines be grown on a smaller scale before aseptic transfer to the entire heat exchanger takes place.

One advantage of such a method is that a subculture that can be grown on a smaller area Can, first of all, be tested for changes in morphology, karyology, and lifespan before cultures become larger Extensive to be created. If the subculture is mutated or contaminated, it can be abandoned. j

The selection can be made with the help of a connection network.

In one embodiment, the rate of the vaccine remains and the growth medium is substantially constant in all of the process flow spaces.

Speed is understood here to mean both the direction and the speed of movement of the flow. It it was found that certain cell lines grow better if the growth medium has a cell monolayer that is attached a vertical surface adheres in an upward direction while all other cell lines flow downward have a flowing median.

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By ensuring that the flow rate is substantially constant, the preferred flow for a special cell growth line can be maintained over the entire surface of the plate.

In one embodiment, the effective area on which the cell tissues are grown is determined with the aid of an insert enlarged, which is used in any one or more process flow spaces of the heat exchanger.

By increasing the surface area with the help of inserts you can the total number of cells that can be grown in the heat exchanger can be increased.

The insert can be a polypropylene grid, stainless steel wire mesh, or a perforated metal plate be.

By inserting any of the above mentioned inserts between the plates of the process flow space, the total area, which is available for cultivation is increased by up to 65% with the same internal volume of the heat exchanger will. The polypropylene grille is less expensive than a stainless steel metal grille or plate. One such However, polypropylene mesh is less suitable for use in connection with media that contain solids ; or other interfering components, since it is much more difficult to keep the mesh openings clean and to sterilize. .

The insert can be a plate of a plate heat exchanger!

The effect of adding an inserted plate to the stack is to reduce the surface area of the process flow spaces by 100% or more by adding a number of process flow spaces between each pair of Has flow spaces for an auxiliary liquid.

In one embodiment, the adhesion of the cell tissue to; the surfaces of the flow spaces reinforced by a pretreatment 'of these surfaces. The pretreatment can be done by sand ' blasting or glass blasting the surfaces of the flow spaces! take place. The resulting scar-covered surface j provides a suitable environment for both the initial | Capture as well as good long-term adhesion of these cells', which otherwise do not easily adhere to smooth cells.

The pretreatment can alternatively include the application of a surface coating to the surface of the flow space include.

By covering the surfaces of the flow spaces with, for example Polylysine, histones, protamines, collagen, gelatin, glass or polystyrene plastic with protons that By having the superficial aromatic groups discharged, the affinity of the cells for the surface can be increased will.

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A proteolytic agent such as trypsin or pronase can be used to release the cells from the surface. The advantage of a plate heat exchanger as a culture vessel is that the temperature can easily be lowered to values below 15 ° before the treatment with the proteolytic agent takes place. At a temperature below 15 ⁰ C the access of trypsin and pronase to critical cell components is greatly reduced. In addition, the low temperature reduces the degree of adhesion to the surface at the side corners of the cells and increases the free-!

laying down the remaining areas of liability. These effects j

I cumulate the reduction in both the required trypsin ^ or pronase activation units as well as the exposure time for the release of the cells.

In one embodiment, the cell tissue is adhered to the surfaces of the flow spaces by means of a pretreatment these surfaces diminished.

The pretreatment involves the application of a thin film coating of polytetrafluoroethylene to the surface of the Flow spaces.

This area is suitable for cells that have only one relative require slight adhesion to the surface and which must be easily removed.

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In one embodiment, a duplex seal arrangement
applied.

When the cells or products of the cells of especially
are of a guaranteed nature, the plate heat exchanger in
be constructed of such a shape that there is a leakage
any hazardous material to the atmosphere or
of the temperature control medium through the sealing means
impede. Such an accident-proof seal to prevent any escape of permissible fluids
to the atmosphere or into the auxiliary liquid is in the
British patent application 206 28 33A. A monitoring system is also described there to provide an early warning of any leakage by the first seal
admit. This can be especially important if it is
the cultures are highly pathogenic organisms.

In one embodiment, the decomposition of the j

i Run surface adherent cells in place. j

By dismantling the cells adhering to the surface at j

Place, the content of the cells recovered ι

\ will. The decomposition can be carried out in a variety of ways

follow, e.g. through virual infection, through temperature |

change or by an osmotic fancy. ι

In accordance with a second aspect of the invention j there is provided an apparatus for j

breed. This device comprises a plate heat exchanger, means for circulating a temperature controlling medium through one set of flow spaces of the heat exchanger, means for circulating a growth medium through the others Sets of flow spaces, means for removing gases from the circulating growth space after it passes through the heat exchanger left, means for air injection or

• Oxygen in the circulating medium, means of injection an inoculation medium in the circulating medium as well as an instrumentarium for measuring the working parameters, so as to Control process.

; Under certain circumstances, e.g. when a pathogenic

\ Cell line, for cell lines that have been inoculated with a pathogen! are, or in the presence of a cell line growth

; Poison or a carcinogen, it may be desirable to have one

j to use heat exchangers with adjacent plates, which limit a flow space for the process medium, in pairs according to the circumference and around the circumference of the continuous

openings, which are the channels for forwarding the auxiliary media limit, welded ,, are., while the flow spaces for the auxiliary media are sealed by flexible seals will.

• In this way use is made of relatively large ones Surface areas such as those used in a plate heat exchanger

; To be available. It can be seen that by adding or Removing panels from the available area easily

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can be adapted to the desired purpose. In addition, \

' by circulating controlled temperature heating or j

'' KUkungsmedien in the interstices the cultivation temperature j

can be set precisely. If a corrugated surface is J

; is applied, the originally introduced cells without j

' ^: the need to reorient the surfaces on which j

the cells to be grown will settle. There is only one

small part of the total growth medium in contact with
the cells at any given time can be a control
j of the process by treating the medium outside of the

j heat exchanger can be obtained by e.g.

! i

; the nutrient content, the dissolved oxygen content and the like.

monitors and regulates. ;

The invention is explained in more detail below with reference to schematic drawings using an exemplary embodiment.

Show it:

Figure 1 is a schematic diagram of a device for
Culturing cells for carrying out the method according to the invention.

FIG. 2 a schematic view of a modified device for growing cell cultures,

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Figure 3 is an exploded view of a plate heat exchanger and

FIG. 4 shows two possible uses in a scheraatic diagram, through which the surface available for cell growth can be increased.

In Fig. 1, one embodiment of an inspection to obtain a monolayer tissue culture is shown, namely below Use of a plate heat exchanger 1A and 1B as a biological reactor with a large surface area. Like it at a plate heat exchanger 1A and TB is normal, this contains a stack of e.g. corrugated sheets (not shown) which are in a predetermined face-to-face relationship. To ensure that the cells are anchored, the plates have contact points between adjacent plates. These can be formed either by crisscrossing and abutting corrugations in the plates, or by locally projections on the corrugations or the like. Provided, which normally ensure the mutual support of the plates. These contact points provide a secure anchorage from which the cells spread over the plate surface can spread as they grow.

A storage tank 2 for a nourishing growth medium with an outlet 3 is connected to a pump A which has the Directs medium from tank 2 via a set of flow spaces (shown at 43) of heat exchanger 1. From the heat

Exchanger 1 can return the medium via a degassing device 6 and a return line 27 to the tank 2 will.

Between the pump 4 and the heat exchangers 1A and 1B is a Injection line 8 is provided which can be used to introduce air or oxygen into the circuit. A discharge line 28 is provided downstream of the pump 4. A storage tank 5 is parallel to the tank 2 in order to circulate a second medium by the pump 4 through the heat exchangers 1A and 1B permit. The medium is fed to the tank 5 via the line 42.

Steam inlets 9 to 16 are provided to be in place and Place to be able to steam sterilize the system. These include the condensate lines 17 to 26. The The degassing device 6 can be designed, for example, in the form of a vacuum vessel. It serves to air and / or To remove carbon dioxide and / or other gases from the circulating medium, thereby avoiding aerobic bacterial contamination to avoid. At the same time, waste products from cell growth can also be removed within the heat exchanger 1A and 1B are removed. The air or the extracted carbon dioxide gas can pass through a sterilization filter 7 be removed, so that it is prevented that pathogenic ι organisms get into the environment.

If no degassing device is required, the excess air can be removed from the injection point 9 simply by degassing.

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, ventilate the tank 2 via a filter 29 can be derived.

The instrumentation for controlling the process includes a pH measuring device 30, a pressure measuring device 31 and a temperature measuring, device 32. The pH measuring device 30 controls the injection of acids or alkalis used for correction via a Line 33. The pressure measuring device 31 controls a pressure valve 34.

The flow spaces that remain between those occupied by the circulating medium are connected to a hot water or cooling water circuit to which a pump 35 belongs. The water can, for example, through a heat exchanger! 36 where it is heated by steam. This .' is fed via a line 37 and a valve 38 which is controlled by the temperature measuring device 32. If cooling is desired, fresh or self-cooled water can be made available via a line 39, which is pumped through the heat exchanger and then discharged through a drain valve 40.

Before starting operation, the device shown must be sterilized. This can either be done by chemical sterilization! medium or preferably by steam at a temperature of 121 ^° C. and for a duration of 30 minutes. The steam inlets 9 to 16 are used for this purpose.

A sterile growth medium containing the appropriate vaccines for the cell line in question can then be found on

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be introduced into the tank 2 aseptically. A sufficient quantity of this inoculant can be put into the plate heat exchanger : Section 1A introduced to the space of the plate; Fill the heat exchanger 1A and the plate surfaces be- j

; j

: rich to flood. Alternatively, a sterile ' Growth medium can be introduced from tank 2, the

; Inoculation with the corresponding cell line takes place in that the inoculum is introduced at the injection opening 44

will. Sterile air or oxygen can be passed through the vaccine

I will be bubbled through by injector 8, if so

j is required. Alternatively, the vaccine material

j also included in both sections of the heat exchanger TA and 1B.

i will be brought. Due to the corrugated nature of the panels

! the cells of the vaccine settle naturally

! on the horizontal components of the panel surface.

! So there is no reorientation or cell attachment mechanism.

I must be necessary, as is notj with flat plates or disks can be agile. The multiple contacts between the plates J or the projections provide a secure anchorage, from I which the cells are made up of via the plate surfaces

ί can spread. In any embodiment that includes an insert j encompassed in one or more flow spaces, the cells will also attach themselves to the surfaces of the inserts. j

After the cell adhesion has been achieved, the inoculation medium can be used

; subtracted from. The system is then again with fresh

j Load the medium, which is then passed through sections 1A or ^! the sections 1A + 1B is recirculated so that the adhering

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Cells can grow to the required density. Air or oxygen can be injected through the opening 8 if necessary. The device 6 for removing air, or CO _2, can be activated to prevent aerobic bacterial contamination and to remove the waste products of cell growth. The medium is then returned to tank 2.

During the cell growth phase, the conditions of the cultures must be kept at an optimal level. The sterile Air that is introduced via injector 8 must be precisely controlled, as must the pH of the medium. the Air or the oxygen are dissolved in the circulating medium. A very precise temperature control can by Circulation of hot or cold water on the service side 41 of the plates using a pump 35 is obtained will.

If only the section 1A as the inoculation section of the plate heat exchanger is used, the cells can be removed when the cell growth has ended by removing the growth medium and filling the system with trypsin solution, by which the cells are detached from the plate surfaces. You can increase the flow rate by To maintain turbulence between the flow plates of section 1A so as to detach the cells from the plate surfaces The cell suspension can then be introduced into both sections 1A and 1B of the plate heat exchanger so as to the cells by infusing them all over the

Available areas of the panels. After the cells adhere to the plates again, the trypsin ; solution to be withdrawn. The system can then be refilled with a growth medium. The procedure is then repeated

¹ fetches until cell growth is complete. It is then raögi borrowed to initiate any further triggering means, e.g.

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¹ a virus or an additive such as sodium butyrate to reduce the |

To stimulate interferon production. !

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¹ Removal of the extracellular products is achieved by completely withdrawing the medium from the system.

^: If the cells are to be harvested by themselves, the same · I

: i

Procedure carried out as described above and finally the '

■ Cell suspension finally discharged via line 28.

i An alternative to injecting air or oxygen into the ; Ventilation purposes consists in adding oxygen to the medium

! oversaturate, such as that in UK patent application 203 47 50A

■ is described.

In FIG. 2, a further embodiment of the device for growing cells ¹ using a plate heat, exchanger 50 is shown.

A storage container 51 for a growth medium is provided with a stirring device 52, a temperature sensor 53 and a sensor 54 for the pH value and can be cleaned with the help of steam

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injection through the opening 56 are sterilized. That Growth medium can be fed to the container through opening 55. Corrections of pH and others Size can be done via an additional opening 74,

A three-way valve 57 connects the reservoir 51 to an inlet port 58 _; a harvesting solution and a pump. This pump circulates the growth medium from the storage container past the inlet and outlet openings 60 into the process flow spaces 61 of the heat exchanger. The growth medium is returned to the reservoir via a flow passage.

The head space of the container 51 can be passed through a conduit 63 via a, - flow measuring device 64 in a metered manner be fumigated. A sterilization filter 65 can be placed in the gas supply line 63 should be inserted. Samples can be drawn off at a sampling opening 70.

The process flow spaces of the heat exchanger are sterilized by steam, which is supplied through an opening 68 and is discharged as condensate through an opening 69.

A vaccine is normally introduced into the system through port 60 to clear the process flow spaces 61 of the Heat exchanger 50 to fill. The temperature of the process and the auxiliary liquids can be measured by sensors 66 and 67. Wake up, excess gases in the system can through

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the sterilization filter 71 can be drained. The auxiliary liquid circulates in the flow spaces 72 and can j

cooled or heated by a unit 73. j

The benefits of both of these systems include:

1. A broader surface for cell growth

2 small volume, up to a maximum of 1500 m.

2. Corrugated surfaces provide good cell adhesion.

3. The area size can easily be changed by removing and adding plates.

A. Good procedural control is possible because of the low filling volume for the medium and the recirculation flow.

5. A rapid equalization of the temperature allows an optimization of the product formation and a minimization of the Cell losses during the transfer between cultivation and Stage of production.

6. Aseptic growth and product recovery are ensured.

Reference is now made to FIG. 3. This shows in FIG exploded view of a plate heat exchanger. In this case, the plates 80 and 84 are between a head and a follow-up piece 82 compressed. A fitting 83 can be used to control the flow of process fluid and the auxiliary liquid via plates 80 and 84 zu.separate, effectively two heat exchangers within of the same frame come to rest.

Using a substantially similar arrangement, the flow of nutrient through section 80 (A1 in Fig. 1), as indicated by dashed lines in Fig. 3, from the flow of the nutrient through the section 84 (IB in Fig. 1), as indicated by a solid line in Fig. 1, are separated, while the auxiliary liquid, which is shown in dash-dotted lines flows through both sections 80 and 84.

Figure 4 shows some possible inserts that can be inserted between the plates of the process flow spaces, in order to be able to enlarge the area required for cell growth.

The perforated metal plate 90 has openings 91 into which the contact points of the adjacent plates, which delimit the flow spaces and intervene. The plate itself can for example be made of stainless steel, titanium or other suitable metal.

The grid 92 may be constructed from a stainless steel wire mesh-j

wire or made of polypropylene. The Cells removed from plates and / or inserts in the manner previously described and from the system washed out. This is followed by cleaning and sterilization using steam. Burning is avoided in this way.

In order to improve the understanding of the invention the following examples are given.

example 1

The device described above has one available

2 standing area for the cell cultures of 2 m_, which by 24 stainless steel plates is formed. This arrangement was used to grow human fibroblasts as follows:

The heat exchanger and the parts of the associated pipework that make up the heat exchanger with the storage tank for the Growing medium links were made by direct injection

2 of steam at a pressure of 1.05 kg / cm for the duration of Sterilized for 60 minutes. The container kettle, consisting of a fermentation kettle made of glass and stainless steel with a j Capacity of 3 liters, which is used together with the rest of the pipes and the pump to circulate the growth medium, was by an autoclave process for a duration of 140 min.

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sterilized at a pressure of 1.05 kg / cm.

Cold water was passed through the heat exchanger the jacket side is cooled down. Then the temperature was im

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Range of 36-37 C by circulating controlled water Temperature stabilized by the shell side. The reservoir, the rest of the piping and the pump were then aseptically connected to the heat exchanger.

An inoculant of 1.8x10 MRC from human fibroblast cells, which in 2.7 ltr. Growth medium were suspended, was then introduced up into the cell culture chamber of the heat exchanger until the chamber was completely filled. The growth medium used was Eagle's medium, which It contains Earle's salts, and was made with 8% ox serum, not essential amino acids and penicillin / streptomycin supplements. The medium was buffered with 2.2g sodium bicarbonate per liter, superimposed with 5% carbon dioxide in air and contained phenor red as a pH indicator.

Additional 3.1 ltr. Growth media was aseptically introduced into the reservoir. The contents were stirred, the head space of the container with 5% carbon dioxide in air gassed j and the contents at 37 ⁰ C and heated thereon; ' Temperature held.

The system was then left undisturbed at 3 ° C. for a period of j

! Rest for 24 hours so that the cells are on the surface of the!

Could attach heat exchanger plates. \

At the end of the sticking period, the heat growth agent in i

the circuit that circulates the reservoir, the pump that:

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The culture side of the heat exchanger contained, at a rate of 100-130 ml per min. For a period of 7 days, such that the flow is up through the heat exchanger
while at the same time water is passed through the shell side

; became. The growth medium was then completely removed from the
Device removed and 5.8 ltr. fresh growth medium
introduced aseptically. The fresh growth medium was with
a speed of 130-280 ml per minute for the duration ■;

, circulated for another three days.

ι:

j The circulation of the growth medium was then stopped.

,

Saline solution (31tr.) Buffered with phosphate was introduced;

'to bring the growth medium down through the heat exchanger _; ■ displace. The phosphate buffered saline solution was then ·. again by 0.25% trypsin in phosphate-buffered saline _; solution (1 liter) replaced. The cell culture chamber has been completely 'deflated and maintained at a temperature of 36-37 ° C, and' \ namely by water directed through the shell side. This happened for a period of 1 hour. Three aliquot parts (21 tr.)
of serum-free growth medium were then sequentially passed up into the cell culture chamber, with a mixture of: 5% carbon dioxide in air pulsing through each aliquot
was withdrawn from the chamber before the filling and the
next aliquot has been added.

i The number of cells that were inoculated and harvested was _:

determined by a Coulter counter and with a hemocyte; meter checked. . i

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Total of inoculated cells 1.8x10

harvested cells

1) 2.2x10 in the withdrawn growth medium

2) in the buffered saline solution and the trypsin solution 8.7x10

3) pulsed serum-free medium 29.1x10

a total of 29.2x10

Example 2

Using the method according to claim 1, human,

I grew fibroblasts as follows: ι

A heat exchanger with 24 plate stainless steel and
an area available for cell culture of
2m x was 1.35x10 MRC 5 human fibroblast cells
vaccinated. The cells were introduced as a suspension in the growth medium, which had the minimum necessary medium
(Eagle) covered with Earle's salts and with 8% ox serum,
non-essential amino acids, penicillin / streptomycin and ■ 2.2g / ltr. Sodium bicarbonate had been supplemented. ί

ι The inoculated container was at 35 ° C for the duration of I

Held like this for 17.5 hours. The volume of the growth medium was!

enlarged to 5.8 ltr. and the growth medium then became!

through the cell culture chamber at a rate of; 100-200 ml per minute for a period of 5 days. The ·

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Flow was through the heat exchanger from bottom to top, and in countercurrent to the flow of water through the shell side. The growth medium was drawn off and replaced with 5.8 ltr. fresh growth medium that was circulated for an additional 5 days. The circulating medium was maintained at a temperature of 34-36 ⁰ C and by passing 5% ι carbon dioxide in air through the headspace of the reservoir with oxygen loaded} pH of the growth media was maintained by addition of sodium bicarbonate at the desired value. ;

The growth medium taken from the cell culture chambers was supplemented with phosphate-buffered saline and the cells for removal from the growth areas by washing with 0.25% trypsin dissolved in phosphate} buffered saline solution. After all of the solutions were withdrawn from the cell growth chamber, the cells were kept at 35-37 ° C for 1.5 hours in the presence of residual trypsin solution. The cells were from the Set up with three aliquots of a serum-free

The growth medium is removed, which is obtained by the injection of Air containing 5% carbon dioxide was stirred while the aliquots were kept in the cell culture chamber became.

Total number of cells vaccinated 1.35x10

harvested cells

1) 1.2x10 ° in the withdrawn growth medium

2) in the buffered laundry solution

and 1.5x10 in the trypsin solution

3) in the pulsed serum-free

Medium 16.1x10 °

a total of 16.2x10

Other checking devices such as analyzers for glucose, dissolved oxygen and carbon dioxide can be used be used.

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Claims (1)

57-72 PATENT LAWYERS \ DR.-ING. R. DÖRING DIPL.-PHYS. DR. J. FRICKE BRAUNSCHWEIG MUNICH Expectations 1. A method for growing cell tissue, thereby g e k e η η draws that you can settle the cells on plates of a plate heat exchanger and a growth medium in continuous circulation through the between allows defined flow spaces to pass through the plates. 2. The method nacji claim 1, characterized in that a virus or other additive is added to the circulating growth medium during circulation. 3. The method according to claim 1 or 2, characterized in that air or oxygen in the circulating Growth medium-is dissolved upon entry into the heat exchanger. 4. The method according to claim 1, 2 or 3, characterized in that g e k e η η ■ indicates that steps are provided to remove atmospheric oxygen or carbon dioxide from the growth medium before leaving the heat exchanger. L J 5. The method according to any one of the preceding claims, characterized in that only ab- ι changing flow spaces for breeding and the da- ' intermediate flow spaces for the circulation of Water or other medium when controlled; Temperature can be used. 6. The method according to any one of the preceding claims, characterized in that a j chose variation in the effective area on which i animal cell tissues are grown, thereby preserved! ι be that you have the inoculum and the growth medium! through selected flow spaces of the heat exchanger by i directs. 7t method according to any one of the preceding claims, characterized in that the speed of the inoculation culture and the growth medium essentially is kept constant in all flow spaces. 8. The method according to any one of the preceding claims, characterized in that the effective Area on which the cell tissues are grown by inserting an insert in any one or more Process flow spaces of the heat exchanger is enlarged. 9. The method according to any one of the preceding claims, characterized in that the adhesion the cell tissue on the surfaces of the process flow spaces is reinforced by a pretreatment of these surfaces. 10. The method according to any one of claims 1 to 8, characterized in that the adhesion of the Zeil-; tissue on the surfaces of the process flow space is reduced by pretreating these surfaces. 11. The method according to any one of the preceding claims, characterized in that a double seal is used. 12. The method according to any one of the preceding ^claims: characterized in that one dismantles: the cell adhering to the surface in place executes. 13. The method according to any one of the preceding claims, characterized in that the composition the inoculation medium and the growth medium, the temperatures of the process and the service fluid, and the flow rate is monitored and controlled with the aid of electronic control means. 14. Device for growing cell tissue, in particular for carrying out the method according to one of the preceding claims, characterized by a plate heat exchanger, means for circulating a temperature control medium through a set of flow spaces of the heat exchanger, means for circulating a growth frediura through the other set of flow spaces, means for removing gases from the circulating growth medium, after it has left the heat exchanger, means for injecting air or oxygen into the circulating one Growth medium, as well as means for injecting an inoculating medium into the circulating medium and through a Instrument device for measuring the working parameters to control the process.