TWI250207B - Double-chamber bioreactor for tissue engineering - Google Patents

Abstract

A double-chamber bioreactor for tissue engineering capable of cultivating two types of tissue or cell at the same time includes: two containers each having an opening, and a perforated dividing plate connecting the openings of the containers and equally dividing the containers into two separate chambers. The perforations on the dividing plate are used to support a carrier for adhesion and growth of cells thereon. The liquids contained in the chambers do not intermix with each other and are separated from each other by the dividing plate so that the two sides of the carrier are separately located in different chambers. Two tissues with a 3D structure can be simultaneously cultivated on a same carrier after each chamber is separately loaded with a required culture solution.

Description

1250207 V. INSTRUCTION DESCRIPTION OF THE INVENTION (1) Technical Field, Field of the Invention The present invention relates to a tissue engineering dual-chamber bioreactor for culturing a tissue having a 3D structure, and more particularly to a culture for at least two The seed cells are grown on the same carrier into two different sets of bio-reactors. The disadvantage of the prior art body age gradually changing the physiology to replace the filling to make it different from the common implants is that the full replacement of the missing body is different after the body is replaced. Common species repels the reaction, but suffers from it, so it can be organized or mechanized to produce functions. The damaged part can be non-biological, such as metal joints, etc. The organization of such materials or the method can make the wear and tear solution can be distinguished. The example of 楂 is #1 is because the energy transfer will often cause defects in the patient. It can be made only by one-way nature and life, using the normal organs, and the period of use is implanted as "the patient's skin transplant system suffers from the damage of the disease itself. The artificial abiotics are moved according to the physical properties. There is no further, 'so the implant, the biological body transplanted on the normal planting. From the original caused by the second dye, the external force of the tissue or the surrogate (the transplanted body implants two kinds of bones, high-material materials transfer biological activity Materials patients need to cause the patient's transplant," and "different parts of the body transplant, although the health damage. Damage or with the organ damage to no called the transplant body" material, can be. Abiotic transfer molecules Polyethylene is implanted into the body's largest 'so it is impossible to complete (such as polyethylene) planting surgery for a great inconvenience. According to the transplant body, the two parts of the damaged part can avoid the poor condition of the disease and the body Transplantation

1250207 V. Description of invention (2) Must wait for others to donate and control the possibility of reducing the immune response to other diseases after being implanted in the body. There are many, but there is no demand at present. If you can make a living, it will be a great boon. Tissue engineering is an exhibition. Its purpose is to hope that a functional organization or device is a substitute for aging and defects. It is necessary to cultivate suitable conditions: first, there must be a carrier (Scaffold), and the growth and development needs It can be placed in the environment of general tissue growth, etc., so that the tissue can be woven in a fast growing environment. It can be known from the above that the cultivation can be carried out smoothly. It consists of a kind of tissue, and a tissue system. Therefore, if there is a suitable organ in a body with a 3D structure, even if there is a suitable organ, a serious rejection phenomenon for the epidemic reaction is necessary, but it may increase the patient. It can be seen from this that although patients who need to be transplanted have a complete transplant system that can cope with the artificial organs that need to be implanted in the body, the patient has a new official, and the organ transplantation is suitable for subdivision and culture. In contrast to our group and the outside of the group, the cell culture of Shengxing is used to continue the growth of the cells in the body. The jfflL must be long-haired, and the device should be known to the research. Internal repair execution function. The internal tissue or the cell and the factor that can be inserted into the vector. In vitro culture, but the necessary nutrients and gas sports, and responsible for mentioning (the environment provided by Bioreactor, the majority of the human body has been producing a damaged tissue or tissue engineering in vitro. Attachment must be provided when the culture group is required to provide physical and woven materials. A suitable stimulating cell group determines that the organization's officers are not only one but also have a certain degree of linkage in the transplant body and have a single culture group to raise the direct response. Can't only

Page 6 1250207 V. INSTRUCTIONS (3) The function of weaving' must have the ability to culture two or more cells or tissues at the same time. At present, there are several bioreactors commonly used to culture tissues or cells. (1) Hollow fiber bioreactor (H〇; [i〇w fiber bioreactor) (2) Spinner nask (3) Rotary volume Rotation vessel (4) Perfusi〇n chamber 〇 Hollow fiber bioreactor, also known as Artificial Capillaries Reactor, is the most common bioreactor at present. Such a bioreactor has been disclosed in U. The disadvantage of this type of reactor is that cells can only grow in the interstices between the fiber tubes, so that tissues having a 3D structure cannot be cultured. Although the other three designs can be used to culture tissues having a 3D structure, none of them can provide two different culture solutions at the same time to simultaneously culture two different tissues on the same carrier. A bioreactor capable of culturing a tissue having a 3D stereostructure has been disclosed in U.S. Patent No. 6,004,088, but it is mainly used to culture liver tissue composed of two kinds of cells on a planar carrier (M esh ) ( A single tissue), its three-dimensional structure comes from the continuous proliferation of cells, but there is no function of culturing both tissues at the same time. In summary, there are currently no methods for culturing two or more different types of tissues or organs at the same time. From the above, if you want to train in vitro

1250207 V. INSTRUCTIONS (4) A bioreactor that can be directly transplanted into the body and has a 3D structure, a bioreactor that can be used to simultaneously culture both tissues is urgently needed. SUMMARY OF THE INVENTION One object of the present invention is to provide a tissue engineered dual chamber bioreactor that can culture tissue having a 3D steric structure. Another object of the present invention is to provide a tissue engineered dual chamber bioreactor that can simultaneously culture two cells or tissues on the same carrier. The additional advantages, objects, and features of the invention will be apparent from the description of the appended claims. The objectives and other advantages of the present invention will be realized and attained by the written description In order to achieve the above objects and other interests and in accordance with the purpose of the present invention, the present invention provides a tissue engineering two-chamber bioreactor comprising first and second containers, each having an open side; The separator partitions the first and second containers, and the two sides of the separator are respectively connected to the open sides of the first and second containers, so that the first and second containers respectively form two independent chambers. The first and second containers further comprise at least one culture fluid injection port; at least one culture fluid discharge port; and at least one gas inlet and outlet. In another aspect, the separator for separating the first and second containers further comprises at least one hole for erecting cells or groups

Page 8 1250207 V. Description of the invention (5) The plant is attached to the growth, the carrier is inserted into the hole, and is partitioned by a partition, and the two ends are suspended in the first container and the second container, respectively. The first container and the second container are formed into two separate closed chambers. The cell line is implanted at both ends of the carrier, and the culture medium required for culturing the first tissue or cell is placed in the first container. The culture required to culture the second tissue or cell is placed in the second volume. The two tissues or cells can be cultured simultaneously on the same carrier. Depending on the needs of the cultured cells, different growth factors can be added at specific time points to promote cell growth and differentiation.

The present invention will be further understood from the following general description and the detailed description of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT The preferred embodiments of the present invention are described in detail with reference to the drawings.

Referring to the first figure, the first figure is a schematic diagram of the tissue engineering dual chamber bioreactor of the present invention. As shown in the first figure, the tissue engineered dual chamber bioreactor comprises a first container (1) and a second container (2) separated by a partition (3). The first container (1) and the second container (2) each have an open side. Further, each of the two containers is provided with a culture solution injection port (1 1 ) (2 1 ), a culture solution discharge port (12) (22), and a gas inlet and outlet port (13) (23). a partition (3) disposed between the first container (1) and the second container (2), the two sides of which are respectively associated with the first volume

Page 9 1250207 V. Description of the invention (6) The open side of the device (1) and the second container (2) are connected, and the container and the partition are locked by a fixing clip (5) to make the first container (1) And the second container (2) respectively form two closed chambers. The air in the two chambers can be replaced by the gas inlet and outlet (1 3 ) ( 2 3 ), respectively. The culture solution can be separately injected into the two chambers through the culture solution injection port (1 1 ) ( 2 1 ), and discharged from the culture solution discharge port (1 2 ) ( 2 2 ), respectively. To avoid contamination, a filter membrane (not shown) can be added to the gas inlet and outlet (13) (23) to prevent bacterial impurities from entering.

The second figure is a plan view of the partition (3) of the tissue engineering dual chamber bioreactor of the present invention. As shown in the second figure, a partition (3) disposed between the first container (1) and the second container (2) is provided with at least one hole (3 1 ). The carrier (4) is inserted into the hole (31) and mounted on the partition (3), and both sides of the carrier (4) are partitioned into the two separate chambers. In the above separator (3), the number of holes (31) may vary depending on the number of carriers (4) to be erected, but the number of holes (31) is preferably the same as the number of carriers (4) the same. It is also possible to match the size of the desired graft size adjustment carrier and the hole in which the carrier is placed on the silicone spacer. However, the radius of the carrier (4) is preferably slightly smaller than the radius of the hole (31) so that the carrier (4) can be inserted into the hole (3 1 ) without passing the culture liquid phase in the two chambers.

In the tissue engineering double-cavity bioreactor of the above first figure, the outer periphery of the carrier (4) may be wrapped with a stainless steel mesh and then fixed on the separator (3) by using hot melt glue. At this time, both ends of the carrier (4) are respectively located on both sides of the spacer (3). The separator (3) and the carrier (4) adhered thereto were immersed in 75% alcohol for 15 minutes and then taken out. After drying in a sterile environment,

Page 10 1250207 V. INSTRUCTIONS (7) Cells or tissues to be cultured at the ends of the carrier (4). The slab (3) of the body (4) is placed in the first place. The erecting is carried so that the open side of the two containers is opposite to the #='-valley (2), and the _ Ο Ο Ο / / 对 对 对 对 对 对 对 对 对 对 对 对 对The two valleys () are fixed (1) (2) to the partition (3) and locked. The material of the carrier (4) is a scorpion-shaped makeup, @^ 1 糸 , , /, 仏 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞 细胞Since the cultured tissue is to be directly transplanted into the body, it is preferable to use a material having both bio-grain and biodegradability, such as collagen, hyaluronic acid/polylactic acid (polylactic acid). Acid, PLA), p〇iyglycolic acid (PGA), a copolymer of PLA and PGA (PLGA), and the like. Then, in the two separate containers, the culture medium required for the growth of cells or tissues injected into the carrier (4) from the culture solution is injected into the medium (11) (2 1 ), which may be the same or different culture medium. . Then, the injection port and the discharge port of the culture solution are closed, and the tissue engineering double-chamber bioreactor is moved into an environment suitable for cell or tissue growth (such as a carbon dioxide incubator) to culture the cells or tissues of the carrier to be implanted into the tissue. After growing for a period of time in the engineering double-chamber bioreactor, the culture solution is discharged from the culture solution discharge port (12) (22) of the two containers, respectively, and the culture solution is discharged into the 'release clamp (5), and the well-known cells or The carrier of the tissue is taken out to obtain a vector in which both cells/tissue are simultaneously implanted. This vector can be used directly as a transplant. As another embodiment of the apparatus of the present invention, a magnetic stirring rod (丨5) (25) may be placed in each of the two containers in the above assembly reaction, and

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1250207 V. Description of invention (8)

The assembled two-piece T (1 4 ) ( 2 4 ). Slow Rotary Stirring Disturbance = Place a magnetic stirrer to ventilate and discharge the metabolic waste while the nutrient solution is used to facilitate the mechanical stimulation of the cells. It can be used to provide tissue growth. In addition, the culture solution can also be connected to (11) (21) and the culture medium by means of Note 7 (12) (22), respectively. The cavity & pu (not shown) forms a two-loop for automation, and can also connect the temperature, gas pressure, and control enthalpy of the culture solution in a reactor. The control unit (not shown) measures glucose or cells in the culture solution ^ Set f and gas circulation, and borrow the growth situation. By changing the content of the cultivating waste, the different mechanical thorns required for the culturing culture can be monitored. The direction of the entry and exit can also be provided on the side (the original culture solution discharge port, for example, when the culture solution is modified), and the tissue can be provided. When the pressure is 22^ (the direct flow meter culture liquid flows from the side of the carrier. The right side is intended to provide shearing force, then the tissue of the present invention is cultured on the body of the two cells 戋g = reactor, except for Injecting the two sides of the container with the differentiation energy carrier | and injecting the desired induction factor em 2 into the two containers. The tissue engineering male 2 Ϊ::t into different tissues | stretched into three chambers, four chambers or eve The concept of the second person is reversed, and the concept of f can also be considered as needed! The carrier is cultured in the form of a biological reaction state of the species, which is further applied to the households; ΑΓ \, the transplant of four or more tissues. The double-cavity bioreactor is required to discharge the ring system in the body of the double-chamber bioreactor.

1250207 V. INSTRUCTIONS (9) Tissues having a 3D structure are cultured in vitro, and the problem of simultaneously culturing at least two tissues on the same carrier has been solved. The above description is intended to clarify the preferred embodiments of the present invention, and is not intended to limit the scope of the present invention, and all modifications and improvements may be made without departing from the spirit and scope of the invention. Within the scope of the patent of the present invention.

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Claims (1)

125 Mouth 6, patent application scope 1. A bioreactor for culturing cell tissue, comprising two containers with openings for connecting the two separated phases for supporting the fine and forming the mutual holes Inconsistent cavity 2 · such as a 3 · as a 4 · such as chamber nutrient 5 · such as reaction culture 6. such as device 7 · such as room use less two indoor application biological application bio patent compatible patent degradation application patent -^ Steps are all exported. Apply for a patent for further liquid. Applying for a patent is a magnetic patent application to inject cells. The scope of the scope is provided with a partition that ranges from the range of the agitating range to the opening of the container, which can separate the two containers into a chamber, wherein the partition is provided with a hole, which is attached to the cell or tissue. The carrier is such that the chamber liquid separators are demarcated such that the sides of the carrier are in different bioreactors according to item 1, wherein the carrier is supported. The bioreactor of item 1, wherein the carrier is a carrier. The bioreactor according to Item 1, wherein each of the gas inlet and outlet, a culture solution injection port, and the bioreactor of the first item, wherein the organism is at least a stirring device for stirring the chamber The bioreactor according to item 5 of the invention, wherein the stirring device. The bioreactor according to item 1, wherein the medium is cultured such that the carrier can be cultured simultaneously to Page 15