RU2090609C1 - Flow-type biological reactor - Google Patents

Abstract

FIELD: biological engineering. SUBSTANCE: device has reservoir for nutrient medium, closed with air-tight cover and connected by means of flexible tube to reservoir for culturing so that the reservoir for nutrient medium and the connection unit have the property to force out the nutrient medium from the reservoir by heating air in it. The bioreactor can also additionally have reservoir for spent nutrient medium connected by means of tube made of flexible material to the reservoir for culturing. EFFECT: enhanced effectiveness in culturing cells and tissues. 5 cl, 1 dwg

Description

 The invention relates to biotechnology and can be used to cultivate cells and tissues of plant and animal origin, as well as to study the influence of various environmental factors on cultivated objects during biological research.

 A device is known that is widely used in the practice of biological research, which is a flat cup (Petri dish) containing a liquid or hardened (agarized) nutrient medium and closed most often by a lid that is not hermetically attached to the cup walls [1] The main disadvantage of this device is the low protection of cultivated objects from external infection and the need for frequent transplantation (after 1-3 weeks) of these objects to a fresh environment to eliminate their self-poisoning by their vital products, which yazano more each time with an increased risk of introducing external contamination.

 A device is also known for cultivating cells with the circulating movement of the nutrient solution in it (circulating bioreactor), consisting of vessels for the nutrient medium and growing cells, two tubes, each of which connects the bottom of one vessel to the top of the other, and a peristaltic pump (see Agricultural Biotechnology plants / transl. from English M: Agropromizdat, 1987, p. 50-55).

 The latter device is taken as a prototype in connection with the closest closeness in its technical essence to the claimed one. The disadvantage of the prototype is also, although to a lesser extent than in the analogue, the low protection of the cultivated objects from external infection and the possibility of self-poisoning by their vital products that accumulate in a circulating nutrient medium.

 The objective of the invention is to provide a simple and reliable device with a high degree of protection against external biological infection and the removal of harmful waste products from the vessel for the cultivation of biological objects.

 This problem is solved in a device consisting of a vessel for nutrient solution, a vessel for growing biological objects, a connecting tube between them, moreover, according to the invention, the vessel for the nutrient solution is equipped with a hermetically sealed lid, the connecting tube is made of flexible material, the device is also equipped with a vessel for the spent solution when it is tightly connected to a vessel for growing biological objects and the use of a flexible tube for connecting these vessels th material.

 From the given essence of the device it is seen that one of its main differences is the equipment of the vessel for the nutrient solution with a continuous hermetically closing cover. The presence of this structural element eliminates the microbiological contamination of the nutrient solution during operation of the device. At the same time, it becomes impossible to infect a vessel for growing biological objects from a nutrient solution. In the case of infection in the culture vessel, the nutrient medium remains sterile in its vessel, since this solution is separated from the culture vessel by the airspace.

 Another difference is the implementation of the connecting tube between the vessels for the nutrient solution and cultivation of flexible material. This technical solution allows you to install the vessel with the nutrient solution at different heights relative to the vessel for growing biological objects, which gives additional guarantees against uncontrolled outflow of the nutrient medium in the vessel for growing (for example, in the case of sharp daily fluctuations in temperature in the working room).

 Another difference of the device is that it is equipped with a vessel for waste solution, hermetically connected to a vessel for growing biological objects. The introduction of such a structural element provides protection of biological objects from microbiological infection from the side of the spent solution. In addition, the presence of this vessel allows you to accumulate and save in sterile conditions the vital products of cultivated objects.

 And finally, the last difference is the use of a tube of flexible material to connect the culture vessels and the spent solution. This design solution allows you to set the vessel for the spent solution at different heights relative to the vessel for cultivation. This ensures the maintenance of the level and the corresponding necessary object of the nutrient solution in the culture vessel at the level of the outflow point from the connecting tube in the waste solution vessel.

 In addition, the device has a number of other positive properties, which in the first place include: the ability to control the gas environment in the vessel for cultivation; the possibility of replacing the composition of the nutrient solution in the process of cultivating the object without moving it to another vessel and the associated risks of infection and mechanical damage to the object; ease of feeding and discharging solutions into a culture vessel without using mechanical devices (water, air pumps), etc.

 The inventive device is illustrated in the drawing, which shows: valve 1 for the discharge of the spent solution; waste solution 2; vessel 3 for waste solution; gas exchange tube 4 with a filter on the vessel for the spent solution; substrate 5; vessel 6 for growing biological objects; gas exchange tube 7 with a filter on the vessel for growing biological objects; nutrient solution 8; a vessel 9 for a nutrient solution; cover 10; connecting tube 11; connecting tube 12.

 The operation of the device is illustrated by the following example. According to this example, a clamp type valve 1 is used in the device; for vessel 3, a 100 ml glass container was used and having a hermetically closing lid with openings for a gas exchange and connecting tube; the gas exchange tube 4 is made of glass and partially filled with cotton wool as a filter; as substrate 5, sand is used; for the vessel 6, a 100 ml glass container was used, which also has a hermetically closing lid, which is also hermetically connected to the gas exchange and connecting tubes; gas exchange tube 7 is similar to tube 4; for vessel 9, a 300 ml narrow-necked glass container was used; as cover 10, a solid rubber stopper was used; the connecting tube 11 has a diameter of 5 mm and a length of 150 mm and is made of rubber; the connecting tube 12 has a diameter of 10 mm and a length of 30 mm.

 Work with the device begins with the sterilization of its main parts and the planting of the cultivated object on a substrate. Sterilization and landing are carried out according to generally accepted methods.

 After planting, the vessels are connected to each other under sterile conditions (in a laminarbox) as a whole, as shown in Fig. 1. In this case, the culture vessel is located directly above the waste solution vessel, and the nutrient solution vessel is located below the culture vessel.

 To supply the nutrient solution to the vessel 6, the air in the vessel 9 is heated, as a result of which the air expands and displaces the nutrient solution into the vessel 6. After the supply of the necessary amount of solution, the heating is stopped, as a result of which the air in the vessel 9 is cooled, compressed and sucks the corresponding part of the air from vessel 6.

 Air heating can be done with special heaters located near the upper part of the vessel 9 or inside it. With the small size of the device, as in this example, heating can be done by touching the upper part of the vessel with a hand.

 The waste solution is drained by gravity through the substrate 5 and the connecting tube 12. Since in this example the outflow point is below the bottom of the vessel 6, no free volume of the nutrient solution is formed in this vessel. The required amount of nutrient solution for the life of the cultivated object is in the substrate and held in it by capillary forces.

 When the solution is periodically supplied from the vessel 9, the spent solution in the substrate is replaced with a fresh one. Thus, the flow mode of the solution’s movement in the device is achieved, the removal of harmful vital products of the cultivated object is achieved, and the optimal conditions for its development are maintained.

 If it is necessary to change the gas medium in the vessel 6, a gas of the required composition is supplied to this vessel through the gas exchange tube 7. This gas enters the vessel 6, displaces the air therein through the connecting pipe 12 into the vessel 3, from which excess air is released through the gas exchange pipe 4 .

Claims (5)

 1. Flowing bioreactor, consisting of a vessel for a nutrient solution, a vessel for growing biological objects, a connecting tube between them, characterized in that the vessel for the nutrient solution is equipped with a continuous hermetically closing lid, while the vessel for the nutrient solution and its connection with the vessel for cultivation is made with the possibility of displacing the nutrient solution from the containing vessel by heating the air contained therein.  2. The bioreactor according to claim 1, characterized in that the connecting tube between the vessels is made of flexible material.  3. The bioreactor according to claim 1, characterized in that it is equipped with a vessel for waste solution, hermetically connected to a vessel for growing biological objects.  4. The bioreactor according to claim 3, characterized in that a tube of flexible material is used to connect the vessel for growing biological objects with the vessel for the spent solution.  5. The bioreactor according to claim 1, characterized in that the hand of the user of the bioreactor is used as a heater, applied for this to the upper part of the vessel with the nutrient solution.