CZ284540B6 - Apparatus for irradiating cells - Google Patents

Abstract

Potential transplant cells are irradiated with ultraviolet (UV) light. A cell suspension is fed to one end of a transporting path which guides it around an outer cylinder (22) surrounding the UV tube so that it is exposed to the UV radiation. The appts. is ventilated by blowing air into a space located between the outer surface of the UV tube and an inner surface of an inner cylinder (26) positioned between the outer cylinder (22) and the UV tube. The air then flows via apertures (60) at one end of the inner tube into a space between an inner surface of the outer cylinder (22) and the outer surface of the inner cylinder (26). The air finally exits the appts. through vents (68). The irradiated cell suspension is collected in a reservoir.

Description

The present invention relates to an apparatus for irradiating liquids. In particular, the invention relates to an apparatus by which cells can be irradiated with an ultraviolet light source.

BACKGROUND OF THE INVENTION

Transplantation of cells into allogeneic cells of another recipient has been investigated by a number of medical researchers for the treatment of diseases and diseases. In order to successfully carry out such cell transplantation, it is necessary to suppress the resistance of the antigen and / or to recognize the transplanted cells. To some extent, the body's natural tendency to reject allogeneic cell transplantation can be avoided.

One commonly known method for reducing resistance to antigen and recognizing allogeneic cells is to expose the cells to ultraviolet radiation. The use of ultraviolet radiation in relation to cell transplantation is described in an article by H. Joachim Deeg, entitled "Ultraviolet 20 Irradiation in Transplantation Biology", Vol. 45, No. 5 pp. 845-851, May 1988.

Other articles are known which describe particular ways of exposing transplanted cells to ultraviolet radiation. E.g. in one method, blood dissolved in phosphate solution was placed in a petri dish and exposed to ultraviolet light for 20 minutes. The light source 25 emitting ultraviolet light was placed at a specified distance from the petri dish. However, not all experiments seemed to have been performed with a light source located equidistant from the petri dish. Hardy. M.A., Lau, Η. T., Weber, C., Reemtsma, K .: " Pancreatic Islet Transplantation: Immunoaltemation With Ultraviolet Irradiation ", World Joumal of Surgery, Vol. 8, No. 2, pp. 207-213, April 1984. Hardy, M.A., Lau, Η. T., Reemtsma, K .: "Prolongation of Rat Islet 30 Allografts with the Use of Ultraviolet Irradiation, Without Immunosupression", Transplantation Proceedings, Vol. 16, No. 3, pp. 965-869, June 1984.

In another method, platelets were dissolved in solution, placed in an open petri dish to a depth of 1.5 mm, and exposed to ultraviolet radiation while being subjected to continuous 35 vibration. Slichter, S.J., Deeg, H.J., Kenedy, M.S .: Prevention of Platelet Illoimmunization in Dogs

With Systemic Cyclosporine and by UV-Irradiation or Cyclosporine Loading of Donor Platelets', Blood, Vol. 69, No. 2, pp. 414-418, February 1987.

Further, the method includes placing whole blood that has been diluted with Waymouth mines. medium, 40 in petri dishes in a 1.5 mm layer and irradiating the solution with ultraviolet light for min. Deeg, H.J., Aprile, J., Graham, T.C., Appelbaum, F.R., Storb, R., "Ultraviolet Irradiation of Blood Prevents Transfusion-Induced Sensitization and Marrow Graft Rejection in Dogs", Blood, Vol. 67, No. 2, pp. 537-539, February 1986.

In addition to these articles, many other articles have been published concerning the use of ultraviolet light irradiation of cells. See. eg Lindahl-Kisseling, K., Saffwenberg, J .: "Inability of UV-Irradiated Lymphocytes to Stimulate Allogenic Cells in Mixed Lymphocyte Culture", Int. Sheet. Allergy, Vol. 41, 670-679, 1971; Balsh, JD, Francfort, JW, Prloff, L.

J .: The Influence of Ultraviolet Irradiation on Blood Transfusion Effect, Surgery, pp. 243-249, 50 August 1985; Lau, H., Reemtsma, K., Hardy, M.A .: "Prolongation of Rat Islet Allograft Survival by Direct Ultraviolet Irradiation of the Graft", Science, Vol. 223, pp. 607-609, Feb. 10, 1984.

The reviews of the above articles have shown that the techniques and methods used hitherto in ultraviolet irradiation for transplantation and transfusion have a number of drawbacks. Especially

Commonly used techniques are not unified. The nature of the various techniques used is such that even with respect to each individual test it is difficult to maintain uniformity. E.g. if the cell suspension is placed in a petri dish and then exposed to radiation in different ways, the unity can only be maintained if the distance between the cell suspension and the light source is kept constant. Obviously, these distances depend on the amount of suspension placed in the petri dish and of course the amount of cell suspension is a factor that is difficult to maintain constant. Cells, which are placed and suspended in specified amounts of solution, eventually settle on the bottom of the petri dish. Therefore, the number of cells in the suspension tends to decrease during irradiation.

The problem associated with irradiating cells in petri dishes according to the above methods is that it is difficult to expose all cells in the cell suspension to the same amount of ultraviolet radiation. This is partly due to the cell suspension in the petri dishes somewhat stagnating, that is, the cells in the suspension do not move but rather remain in their mutual position. As a result, cells on the surface of the cell suspension lying closer to the ultraviolet light source are exposed to a different amount of radiation than the underlying cells of the cell solution. Although one of the previously mentioned articles mentions shaking the petri dish during irradiation, this technique does not eliminate the problem completely.

E.g. shaking a petri dish that is not covered to expose the cells to movement results in an increased amount of vaporized cell suspension. On the other hand, covering the petri dish before shaking is not the most effective solution since the material from which the cover is made can affect and substantially reduce the amount of radiation received by the cell suspension. Therefore, complicated calibration is required.

Another drawback associated with the technique commonly used in ultraviolet irradiation for transplant / transfusion is that there is no stability with respect to other factors affecting the irradiation process. E.g. the temperature in the space surrounding the irradiation process can have a significant effect on the irradiation intensity. Therefore, if the ambient temperature is not maintained at a certain level, it is not possible to obtain consistent and reliable results with respect to the gradual irradiation process.

Similarly, during the initial hours of operation, the power of the ultraviolet light source may vary into fluorescent lamp systems. Therefore, cell suspensions irradiated during the initial hours of operation will be exposed to a different amount of radiation than cell suspensions that are irradiated later.

SUMMARY OF THE INVENTION

It follows from the foregoing that it is necessary to provide a cell irradiation apparatus that overcomes the drawbacks associated with the techniques used so far. SUMMARY OF THE INVENTION It is an object of the present invention to provide such an apparatus for irradiating cells with ultraviolet light, which will achieve uniformity in irradiating cells in a cell suspension.

It is a further object of the invention to provide an apparatus for irradiating cells with ultraviolet light which permits stability with respect to the operation of the apparatus. Creating an instrument that fulfills both of the above objectives is desirable because, when the instrument is calibrated to ensure that cells in a certain amount of cell suspension are exposed to a certain amount of ultraviolet radiation for a specified period of time, basically the same results as those achieved when calibrating the instrument.

It is a further object of the invention to provide a simple cell irradiation apparatus which is not expensive to manufacture and easy to operate.

-2IS

It is a further object of the present invention to provide an apparatus for irradiating cells which can prevent the passage of ultraviolet light of a certain wavelength so that cells in the cell suspension are not exposed to such light. This eliminates the potential harmful effects of ultraviolet light of a certain wavelength.

The essence of the invention is that it consists of a source of ultraviolet radiation and an outer cylinder surrounding the source. The apparatus is provided with means surrounding the outer surface of the outer cylinder for guiding the cell suspension for irradiation with a source of ultraviolet radiation. Preferably, the device comprises a hollow tube that is helically wound around the outer surface of the outer cylinder. The hollow tube is made of a material that transmits ultraviolet radiation.

According to another embodiment, the apparatus comprises an inner cylinder that is positioned between the outer cylinder and the ultraviolet radiation source. A first hollow space is formed between the inner surface of the inner cylinder and the outer surface of the ultraviolet radiation source, and a second hollow space is formed between the inner surface of the outer cylinder and the outer surface of the inner cylinder, the inner cylinder being made of ultraviolet radiation transmitting material. preventing the passage of ultraviolet radiation of a predetermined wavelength through the inner cylinder from the ultraviolet radiation source.

According to a preferred embodiment, the ultraviolet barrier means may be formed by a thin film disposed on the outer surface of the inner cylinder, wherein the predetermined wavelength is 200 nm to 280 nm, and may further comprise an optical filter disposed on the outer surface of the inner cylinder. the device may be formed by a thin film formed on the inner surface of the outer cylinder, the predetermined wavelength being 200 nm to 280 nm.

According to another embodiment, the outer cylinder is provided with faces to which the sealing members are attached and the inner cylinder is provided with faces on the sides, the first face abutting the first sealing member and the second face abutting the second sealing member. Openings are spaced along the circumference of the second face of the inner cylinder and spaced apart. The first sealing member comprises means (formed by apertures) for connecting the first cavity between the inner cylinder and the ultraviolet radiation source to the atmosphere, and the second cavity between the outer cylinder and the inner cylinder to the atmosphere.

The apparatus is preferably provided with a system ventilation device to maintain a substantially constant temperature.

BRIEF DESCRIPTION OF THE DRAWINGS

An exemplary embodiment of an apparatus according to the invention is shown in the accompanying drawings, in which Fig. 1 is a front view of a cell irradiation apparatus according to the invention; Fig. 2 is a sectional view 2-2 of a preferred embodiment of the apparatus of Fig. 1; Fig. 3 is a sectional view 3-3 of the embodiment of the apparatus of Fig. 2; Fig. 4 is a right side view of the apparatus of the invention; Fig. 5 is a left side view of a preferred embodiment of the apparatus of the invention; and Fig. 6 is a cross-section similar to that of Fig. 2 but showing a different filter location.

DETAILED DESCRIPTION OF THE INVENTION

The cell irradiation apparatus 20 of Figure 1 consists of a hollow outer cylinder 22 that extends longitudinally. The hollow tube 24 is wound helically around the outer surface of the outer cylinder 22. The helical wound tube 24 extends along substantially the entire longitudinal extent of the outer cylinder 22. In a preferred embodiment, the tube 24 is wound around the outer peripheral surface.

3 to 11 of the first cylinder 22 so that each successive thread of the tube 24 is in close contact with the previous one. Thus, the tight arrangement of the helical coil 24 is achieved.

As shown in Figure 2, the hollow inner cylinder 26 is disposed within the outer cylinder 22. The inner cylinder 26 also extends longitudinally and is substantially parallel to the outer cylinder 22. The inner cylinder 26 is spaced from the outer cylinder 22 so that between the inner cylinder the surface of the outer cylinder 22 and the outer surface of the inner cylinder 26 is the space 28. The ultraviolet light source 34 is located centrally in the apparatus 20. The ultraviolet light source 34 also extends in the longitudinal direction and is substantially parallel to the outer roller 22 and the inner roller 26. Thus, both the outer roller 22 and the inner roller 26 surround the ultraviolet light source 34. The ultraviolet light source 34 is spaced from the inner cylinder 26 so as to create a space 36 between the inner surface 38 of the inner cylinder 26 and the outer surface 40 of the ultraviolet light source 34. The ultraviolet light source 34, the inner roller 26 and the outer roller 22 are all substantially the same length.

In FIG. 3, the apparatus 20 further comprises a first sealing member 42 and a second sealing member 44. The first sealing member 42 serves to seal the first face 46 of the outer cylinder 22, while the second sealing member 44 serves to seal the second face 48 of the outer cylinder 22. the member 42 is positioned relative to the outer cylinder 22 such that the first face 46 of the outer cylinder 22 rests against the inner surface 50 of the first sealing member 42. Similarly, the second sealing member 44 is positioned relative to the outer cylinder 22 so that the inner surface 52 of the second The sealing member 44 rests on the second face 48 of the outer cylinder 22, which lies at the other end of the outer cylinder 22.

It is further evident from FIG. 3 that the inner cylinder 26 has substantially the same length as the outer cylinder 22. Therefore, the inner surface 50 of the first sealing member 42 rests on the first face 54 of the inner cylinder 26, which lies at the first end of the inner cylinder 26. the second face 56 of the inner cylinder 26, located at the other end of the inner cylinder 26, abuts the inner surface 52 of the second sealing member 44.

The support members 58 are rigidly connected to the two sealing members 42, 44. The support members 58 serve to support the apparatus 20 in a horizontal position on a horizontal surface. It should be noted, however, that other forms of supports may be provided as alternatives to the support members 58.

Further cobr. At least one and preferably a plurality of spaced apart apertures 60 extend radially through the inner cylinder 26. The apertures 60 are disposed adjacent the second end 56 of the inner cylinder 26, and preferably the apertures 60 extend along the entire circumference of the inner cylinder 26. 44, with the inner and outer cylinders 26, 22 and the arrangement of apertures 60 passing through the inner cylinder 26, the space 28 between the inner and outer cylinders 26, 22 is fluidly connected to the space 36 between the inner cylinder 26 and the light source 34.

The first sealing member 42, as shown in Figure 4, is preferably provided with a first aperture 62 that extends through the first sealing member 42 and is positioned to connect the space 36 between the inner surface 38 of the inner cylinder 26 and the outer surface 40 of the ultraviolet source 34. lights with atmosphere. Due to the tight arrangement of the first sealing member 42 and the inner and outer cylinders 26, 22, the aperture 62 may be connected to the space 36 between the inner cylinder 26 and the ultraviolet light source 34 and the space 28 between the inner and outer cylinders 26, 22 as described above. a suction channel 64 is located in the first opening 62. The suction channel 64 is designed to be connected to a suitable air source 66, such as a small blower. According to this embodiment, the blower may blow or suck air through suction channel 64 and radially spaced apertures 60 into space 36.

The first sealing member 42 also has at least one, and preferably more, second orifices 68 that extend through the first sealing member 42. The second orifices 68 are positioned such that the space 28 between the inner surface 30 of the outer cylinder 22 and the outer surface 32 of the inner cylinder 26 is connected. with the atmosphere. Again, since the first sealing member 42 is tightly connected to the inner and outer cylinders 26, 22, a series of second apertures 68 can only communicate with the space 28.

-4II

Two further openings 70 are formed in the first sealing member 42 through which two hollow pins 72 protruding from one end of the ultraviolet light source 34 may pass. As seen in FIG. 5, the second sealing member 44 is also provided with two apertures 74 through which two hollow pins 76 extending from the other end of the ultraviolet light source 34 can pass. Thus, a suitable power source (not shown) may be attached to the pins 72, 76 from which power is supplied to the ultraviolet light source 34.

Returning to Fig. 1, one end of the helical coil tube 24, the inlet end, is connected to a suitable container 78 in which the cell suspension to be exposed to radiation from the ultraviolet light source 34 is located. The container 78 may be either a transfusion bag or another type of sterile container that may be equipped with an inlet air filter as desired. A variable speed pump 80 is also provided at the inlet end of the helical coil 24. The variable speed pump 80 serves to move the cell suspension housed in the container 78 by the helical coil 24.

A collecting tank 82 is attached to the opposite end, the outlet end of the helical coil 24, in which a cell suspension is collected which has been irradiated with an ultraviolet radiation source 34.

The outer cylinder 22 and inner cylinder 26 should be made of materials that are suitable for passing and filtering ultraviolet light. The material of which the inner and outer cylinders 26, 22 are made should be capable of transmitting at least 90% of the preferred spectrum of ultraviolet light. One material that has been found to give desirable results is fused silica glass. This material has a number of advantages over other materials since the fused silica glass has greater stability when temperature changes. Materials other than quartz glass may be used, provided that the selected material is capable of forming a lens.

Enlarged parts the fluorescent ultraviolet lamp system emits a combination of U.V.-A, U.V.-B and U.V.-C light and differs in their spectral degradation and intensity. E.g. The U.V.-B tubes emit mostly U.V.B light that falls between 280 to 320 nanometers. However, the same tube also emits to a certain extent also a minimum amount of U.V.A and U.V.-C light.

Preferably, a U.V.-B medium wave fluorescent tube is used as the ultraviolet light source 34 in the present invention. Such a tube is suitable because, as mentioned above, it emits an optimal distribution of ultraviolet light in the range of about 280 nanometers (nm). This range of ultraviolet light has been found to be most suitable for irradiating potential transplanted cells in a cell suspension. However, as previously pointed out, the U.V.-B mid-wavelength fluorescent tube emits minimal but additional amounts of ultraviolet light over a wavelength range outside the range of 280 to 320 nanometers. It has been found that ultraviolet light, which is outside the range of 280 to 320 nanometers, is not quite suitable for irradiating potential transplanted cells.

To prevent unwanted wavelengths from passing, a filter device can be provided relative to the inner and outer rollers 26, 22. The ultraviolet light source 34, which is capable of emitting ultraviolet light having the desired spectrum distribution and range, is relatively well adapted to be coupled to a filter device. The filter device allows the portion of ultraviolet light having the desired wavelength to pass through the inner and outer rollers 26. 22. while filtering out unwanted and potentially harmful portions of ultraviolet light outside the UV-B range before reaching the helical coil tube 24 and the cell suspension contained therein .

One part of the ultraviolet light range that has been found to be undesirable is ultraviolet light in the U.V.-C range. It has been determined that ultraviolet light in the U.V.-C range is not well suited for irradiating cells for its high energy and shortwave properties.

-5II

In one embodiment, the filter device has the shape of a film 90 that is formed on the outer surface 32 of the inner cylinder 26 (see reference numeral 90 in FIG. 2). One film that has been found to be very suitable for preventing or blocking the transmission of high energy, short-wave U.V.-C light, is the "KODACEL" TA 401 film produced by Eastman Kodak Co. It is preferred that the film block the passage of U.V.-C light having a wavelength in the range between about 200 nanometers and 280 nanometers. While the above film is located on the outer peripheral surface 32 of the inner roll 26, it is desirable to coat the inner surface 30 of the outer roll 22 with the same type of film 92 (see reference numeral 92 in FIG. 6). For maximum film stability, the filters are preferably positioned away from the ultraviolet light source 34. It has been found that a film of between 0.10 mm and 0.15 mm thickness provides the desired results, but other thicknesses may be used, depending on the desired results.

While it is desirable that high energy short wave U.V.-C light be filtered to prevent irradiation of the cell suspension with this portion of ultraviolet light, filtering and preventing U.V.-A light from passing through is not so important. This is because U.V.-A light has low energy and a long wavelength of light. As a result, U.V.-A light will not have the same potential harmful effects on the cell suspension as high-energy U.V.-C short wave.

However, in order to ensure that the cell suspension will only be exposed to UV-B light, the filter device may be arranged to prevent UV-A light from passing through the inner and / or outer cylinder 26, 22. One particular filter device that appears to be Highly effective is a nickel-cobalt sulfate filter which can be placed at the above locations to filter UV-A light and still transmit UV-B light. As an alternative, any suitable type of optical filter can be used, since the optical filters have the ability to accomplish the desired task of filtering a portion of the ultraviolet light that the cell suspension is not to be irradiated with. It is also possible to place antireflective coatings on the inner and / or outer rollers 22, 26 in order to improve the passage of ultraviolet light through the rollers 22, 26.

It follows from the above that the filter device can be used to prevent and block the passage of U.V.-A light and U.V.-C light, allowing the passage of U.V.-B light. By selecting the film and / or the optical filter, the irradiated cell suspension flowing in the tube 24 is irradiated only with U.V-B light having the desired wavelength.

The above-described filter device can be used in conjunction with different ultraviolet light sources than those already described. E.g. it may be desirable for various reasons to use an ultraviolet light source that emits primarily U.V.-A light, primarily U.V.-C light, or a combination of both.

The helical hollow tube 24 should be made of a material that is adapted to be readily transmitted by ultraviolet light. One of the materials that has been found to pass well through ultraviolet light is polypropylene. "EXTREL" polypropylene and preferably EX-50 polypropylene, manufactured by Exxon Chemical Co., is most suitable.

One of the advantages of the apparatus according to the invention is that the apparatus 20 is equipped with a ventilation system which maintains a constant temperature in the apparatus 20 during operation. Referring to FIG. 3, the ventilation system operates as follows. A small fan 66, or other suitable air source, blows air through the inlet duct 64 and into the space 36 between the inner surface 38 of the inner cylinder 26 and the outer surface 40 of the ultraviolet lamp 34. The air is pushed towards the other end 56 of the inner cylinder 26; apertures 60 that pass through the second cylinder 26. The air then flows as shown by arrows A towards the apertures 68 that pass through the first sealing member 42. Therefore, it is clear that a continuous air flow passing from the fan 66 or other suitable air source to The space 36, apertures 60, space 28, and out of apertures 68 causes continuous ventilation of the apparatus 20 and ensures that air in the closed system is continuously ventilated.

-6 £ 1

One of the advantages of the ventilation system of the present invention is evident when considering the fact that variations in ambient temperature can result in up to 60% variations in ultraviolet lamp power. If the instrument is first calibrated based on results obtained when operating the instrument under certain ambient conditions, further operation of the instrument at ambient conditions other than those at the first instrument calibration will result in the cell suspension being irradiated with the amount, which differ from the expected amount. The use of a ventilation system ensures that the temperature in the device is maintained at a substantially constant level and the cells in the cell suspension are irradiated at the desired level and with the required amount of ultraviolet light.

It will be appreciated that as an alternative to the fan 66, a suction or vacuum pump can be connected to the inlet duct 64, which continuously ventes the closed system. In this exemplary embodiment, a vacuum pump would draw air into space 28 through apertures 68 and air would flow through apertures 60 that extend through inner cylinder 26, aperture 36, and out through channel 64.

A further advantage associated with the ventilation system consists in the above-mentioned use of filters, or the like, for filtering and preventing the passage of ultraviolet light having a certain wavelength. In this regard, the heat produced by the ultraviolet light source 34 and consequently the temperature in the apparatus 20 can be maintained at a level that is not harmful to the filter surface. To further increase the stability of the filter, the air vents 68 through which the air passes between the outside air and the space 28 between the inner and outer cylinders 22, 26 can be switched through the inlet duct 62. Also the air return can be accelerated using the above vacuum or suction pump. heat transfer from the inner and outer cylinders to prevent damage to the filter surfaces. The return air stream allows the heat generated by the lamp to exit the system more quickly as it is routed out of the system immediately instead of passing through the entire system.

The variable speed pump 80 may be replaced by other devices having a similar function. E.g. a Harvard injection pump or other high pressure pumping device may be used to pump the cell suspension from the container 78 by the helical coil 24 into the recovery tank 82.

The use of an outer cylinder 22 having an outer radius of about 24 mm and a length of about 190 mm, measured between the first and second sealing members 42, 44, in conjunction with the hollow tube 24 having an inner diameter of about 1.02 mm and an outer diameter of 2.16 mm. will allow about 17.0 ml of cell suspension to be placed in tube 24 in the irradiation space of the apparatus. The outer radius of the inner cylinder 26 may be about 16 mm. Also, the distance between the surfaces between the inner and outer rollers 26, 22 and the surfaces between the inner roller 26 and the ultraviolet light source 34 may be about 8 mm. These dimensions and values are given by way of example only and are intended only to illustrate the need to know the dimensions of the outer cylinder 22 and the tube 24. to determine the amount of cell suspension contained in the tube 24 to determine the dose for calibration purposes and to achieve consistent results.

In order to ensure that cells in the cell suspension that are pumped by the helical coil 24 are irradiated with a known determined amount of ultraviolet light, it is important and also essential that the instrument 20 be calibrated. As a first operation, the ultraviolet tube or lamp is allowed to burn continuously for about 100 hours in the calibration process. It has been found that the output of ultraviolet lamps varies most during the first hours of lamp operation. Therefore, if the lamp is allowed to burn continuously for 100 hours, changes in lamp performance will be reduced and substantially eliminated.

After the ultraviolet lamp has burned continuously for 100 hours, its performance must be measured. For this purpose, an integration photometer and a spectral radiometer can be used, and the result is the determination of the individual and total powers of U.V.-A light, U.V.-B light and U.V.-C light. During the measurement of the power of the ultraviolet light source 34, the temperature in the apparatus 20 will be maintained at a substantially constant value throughout the operation by the above-mentioned ventilation system.

-7U

Measurements made by a narrowband detector or sensor and laboratory radiometer can be standardized into calibration measurements and used later to determine the variation value of the instrument 20 after, e.g., 6 to 8 months of use. For example, say the calibration measurements taken from the power of the irradiation system are initially 3.0 watts / m ² / s, using a narrowband detector and a laboratory radiometer, the output data can be standardized for calibration measurements.

By using calibration irradiation measurements, the total light output U.V.-B can be obtained from an ultraviolet light source. This power in watts per second can be converted into watts per square meter per second by dividing the surface area of the outer cylinder, or in other words, the area of radiation. Obviously, the irradiation area can be increased by using an outer cylinder having a larger diameter. By adding a film layer such as the previously mentioned film "KODACEL" TA 401 with a maximum thickness of 0.15 mm, undesirable U.V.-C light will be sufficiently filtered. By increasing or doubling the maximum thickness, the amount of U.V.-B light passing through the cylinders and into the cells in the helical coil 24 would be reduced by about 50%. Also the amount of irradiation can be further reduced by controlling the amount of current supplied to the ultraviolet lamp. An adjustable rheostat or a variable energy source can be used to achieve such radiation reduction.

Since the following relationship is known, the pumping rate necessary to thoroughly irradiate cells in the cell suspension can be determined.

dose (J / m ² ) = irradiation (W / m ² / s) x time (s);

where the irradiation is measured after application of the filters and after reduction of the lamp power, if the number of cells needed for transplantation

V (ml) = -------------------------------------- cell number / ml cell suspension where V = volume of cell suspension needed.

V (ml) pumping speed (ml / s) = ---------- time (s)

By way of example, in a typical bone marrow transplantation, the concentration of cells required to rescue an allogeneic recipient was determined to be about 2.3 χ 10 ⁹ cells for 10 rats. Using an outer and inner cylinder having the above dimensions, using a second 0.15 mm thick "KODACEL" TA 401 coating on the outer surface of the inner cylinder (which further reduces ultraviolet lamp power by 50%), adjusting ultraviolet lamp power to 30% and determining For example, if a reasonable dose is about 150 J / m ² , it can be determined from the above relationship that a reasonable pumping rate should be about 2.26 ml / min. when the cell concentration is about 2.3 χ 10 ⁸ cells / ml. .

After determining the appropriate pumping rate, the variable speed pump 80, or other suitable device, is set to this speed and the cell suspension present in the reservoir 78 is allowed to flow to the inlet end of the tube 24. By the pumping effect of the pump 80 and an irradiation surface, wherein the cells in the cell suspension are irradiated with an ultraviolet light source. The cell suspension flows through the entire length of the tube and the entire irradiation surface so as to be exposed to a predetermined appropriate dose of irradiation with ultraviolet light. The irradiated cells in the cell suspension leave the tube 24 at the outlet end and collect in a collection tank 82. The cells are then transplanted into a recipient.

While the foregoing description of the apparatus and method of the invention has been explained by terms used in connection with bone marrow transplantation, it is understood that the method and apparatus can be used in other methods relating to cell transplantation and transfusions.

-81.1

The principles, exemplary embodiments and methods of operating the present invention have been described in the above description. However, the invention to be protected is not limited to the construction of the described embodiment. Furthermore, the embodiments described herein must be considered illustrative, not limiting. Still other changes and variations can be made without changing the spirit of the invention.

PATENT CLAIMS

Claims (10)

An apparatus for irradiating cells with ultraviolet radiation, comprising an ultraviolet radiation source (34), an outer cylinder (22) surrounding the ultraviolet radiation source (34), and means surrounding the outer surface of the outer cylinder (22) for guiding the cell suspension for irradiation with an ultraviolet radiation source (34). Apparatus according to claim 1, characterized in that the means for guiding the cell suspension are formed by a hollow tube (24) helically wound around the outer surface of the outer cylinder (22), the hollow tube (24) being made of a material that transmits ultraviolet radiation. . Apparatus according to claim 2, characterized in that an inner cylinder (26) surrounding the ultraviolet radiation source (34) is disposed between the outer cylinder (22) and the ultraviolet radiation source (34), and between the inner surface (38) of the inner cylinder (38). 26) and a first hollow space (36) is formed by the outer surface (40) of the ultraviolet radiation source (34) and a second hollow space is formed between the inner surface (30) of the outer cylinder (22) and the outer surface (32) of the inner cylinder (26) (28), wherein the inner cylinder (26) is made of a material that transmits ultraviolet radiation and is provided with means for preventing the passage of ultraviolet radiation of a predetermined wavelength through the inner cylinder (26) from the ultraviolet radiation source (34). Apparatus according to claim 3, characterized in that the means for preventing the passage of ultraviolet radiation of a predetermined wavelength through the inner cylinder (26) is formed by a thin film (90) disposed on the outer surface (32) of the inner cylinder (26). the wavelength is 200 nm to 280 nm. Apparatus according to claim 3, characterized in that the means for preventing ultraviolet radiation from passing through the inner cylinder (26) are formed by an optical filter located on the outer surface (32) of the inner cylinder (26). Apparatus according to claim 3, characterized in that the means for preventing the passage of ultraviolet radiation of a predetermined wavelength is a thin film (92) formed on the inner surface (30) of the outer cylinder (22), wherein the predetermined wavelength is 200 nm to 280 nm. Apparatus according to claims 1 to 3, characterized in that an optical filter is arranged on the inner surface (30) of the outer roller (22) to prevent the passage of ultraviolet radiation of a predetermined wavelength through the outer roller (22) from the ultraviolet radiation source (34). Apparatus according to claim 3, characterized in that the outer cylinder (22) is provided on the sides with first and second faces (46, 48), and a first sealing member (42) is attached to the first face (46) of the outer cylinder (22). ) and a second sealing member (44) is attached to the second face (48) of the outer cylinder (22), the inner cylinder (26) is provided with first and second faces (54, 56) on the sides, Wherein the first face (54) of the inner cylinder (26) abuts the first sealing member (42) and the second face (56) of the inner cylinder (26) abuts the second sealing member (44). Apparatus according to claim 8, characterized in that openings (60) are spaced apart from one another at the circumference of the second face (56) of the inner cylinder (26), and the first sealing member (42) comprises means for connecting the first cavity. , between the inner cylinder (26) and the ultraviolet radiation source (34), with the atmosphere, and the second hollow space (28), between the outer cylinder (22) and the inner cylinder (26), with the atmosphere, 36) with the atmosphere comprising at least one first opening (62) extending axially through the first sealing member (42) and adapted to communicate with an air source, the means for connecting the second cavity (28) to the atmosphere comprising at least one second opening (68) extending axially through the first sealing member (42). The apparatus of claim 8, wherein the first sealing member (42) of the first face (46) of the outer cylinder (22) and the second sealing member (44) of the second face (48) of the outer cylinder (22) extends axially at least one first an opening (62) connecting the first hollow space (36) between the inner surface (38) of the inner cylinder (26) and the outer surface (40) of the ultraviolet radiation source (34) with the atmosphere, the first sealing member (42) of the first face (46) of the cylinder (22) extends axially at least one second opening (68) connecting the second hollow space (28) between the inner surface (30) of the outer cylinder (22) and the outer surface (32) of the inner cylinder (26) with the atmosphere; 26) at least one opening (60) extends radially at its second face (56).