TW201546270A - Bioreactor featuring multiple supply flows and method of culturing tissue - Google Patents

Abstract

The present invention provides a bioreactor featuring multiple supply flows, which includes a biological scaffold; a first supply flow section having a flow supply space isolated from the outside to surround periphery of the biological scaffold, a first inflow channel and a first outflow channel each in communication with the flow supply space, so as to enable the flow supply space to communicate with the first inflow channel and the first outflow channel; a second supply flow section having a second inflow channel, which uses one end to communicate with a predetermined space inside the biological scaffold, a second outflow channel having one end in communication with the predetermined space inside the biological scaffold, thereby enabling the predetermined space inside the biological scaffold to communicate with the second inflow channel and second outflow channel.

Description

Multi-feedstream bioreactor and tissue culture method

The present invention relates to biological tissue engineering techniques, and more particularly to a multi-feedstream bioreactor and tissue culture method.

In the research and development of tissue engineering, although the cultivation of specific biological tissues can be carried out through traditional techniques such as hollow fiber bioreactors, it is not ideal for the development of traditional techniques, for the cultivation of tissues in vitro. Usually, only a single culture solution can be provided, so that the culture on the same carrier can only grow a single type of tissue cells, and the application thereof is considerably limited. Then, in the case of the invention patent No. I250207, the focus is on Therefore, instead of the deficiencies of the conventional techniques, a bioreactor having a dual-chamber space is provided, and in each of the two separate chambers, different culture liquids are separately provided, so that different cells grow in the space of the chamber. In this way, the raft is allowed to grow on different cells across a single carrier between the two chambers.

Compared with the conventional technology, the invention patents enable the different cells to adhere to the same carrier, but are limited by the spatial design of the parallel chambers, and different cells are only grown in different positions on the carrier. On the other hand, the difference between the actual biological tissues, such as the liver, the kidneys or the bones, is still too large, and the bioreactors in the two-chamber space cannot be cultured to obtain the tissues and organs for practical clinical use.

Due to the development of medical technology, the average life expectancy of human beings has been significantly extended. In contrast, the proportion of the elderly population will be higher and higher, and in the medical care required by the elderly, the demand for bone treatment due to normal wear, disease or accidental injuries is increasing, especially for large-area bone defects. In the case of tissue engineering, the bone tissue that is sufficient to fill the defect site, supplemented by the intervention of existing medical methods, can promote the rapid recovery of the patient's bone defect tissue, which is a better means for the effective use of medical resources. .

However, as the above-mentioned tissue engineering is limited by the lack of bioreactor technology, when the static culture of three-dimensional tissue is to be performed in vitro, the nutrient transport required for cell growth can only be transmitted to the three-dimensional tissue due to the blood supply to the ischemic tube. In the structure of the project, in addition to thin tissue such as skin or natural ischemic femur such as cartilage, other biological tissues grow more than 1 mm, usually only the surface cells can survive, therefore, in the culture In the development of tissue engineering of larger volume three-dimensional tissue, there is still a lack of bioreactor technology.

Therefore, the main object of the present invention is to provide a multi-feedstream bioreactor for cultivating a bio-scaffold in a tissue engineering development to culture a biomimetic micro-environment in a multi-supply environment. , thereby providing different environments required for tissue growth, so that each supply stream can provide separate exchanges such as nutrient transport, gas and cell tissue growth metabolites, or different operating streams through control of operating conditions. Field and pressure, etc., in response to the needs of the organization of the implementation of the project.

In order to achieve the above object, the multi-supply flow bioreactor provided by the present invention comprises a biological scaffold; a first supply flow portion having a supply flow space surrounding the outside of the environment a first inflow channel and a first outflow channel are respectively connected to the supply flow space, and the supply flow space is connected to the first inflow channel and the first out a second supply flow portion having a second inlet passage connected at one end to a predetermined space inside the biological support, and a second outlet passage connected at one end to a predetermined space inside the biological support And the second inflow channel and the second outflow channel are connected by a predetermined space inside the biological scaffold.

Wherein, the first supply flow portion has a hollow outer cavity member for accommodating the biological support inside the hollow.

The first inflow channel and the first outflow channel are respectively disposed on the outer cavity member.

Wherein, the outer cavity member is tubular.

Wherein, the first supply flow portion further comprises two end pieces, which are respectively disposed at two ends of the outer cavity member tube shaft for closing the inner space of the outer cavity member to be isolated from the outside, according to The supply flow space is formed.

Each of the end members has an end cover, and the cover is disposed at an axially corresponding end of the outer cavity member, so that both ends of the outer tube member shaft are closed by the end caps, and a joint is provided. The outer cavity member is disposed between the biological support and the corresponding end cover.

The second inflow channel and the second outflow channel are respectively disposed on each of the end pieces.

In addition, in order to meet the demand for the supply of the large tissue culture, the number of the second supply flow portion can be two, and the second inlet flow path respectively connected to the different space inside the biological support and the The second outflow channel or the number of the second inflow channel and the second outflow channel are respectively a plurality, and are respectively connected by different spaces inside the biological stent.

Furthermore, in order to facilitate the supply of nutrients in large tissue culture, the present invention further discloses a method of tissue culture, which is a tissue culture using a biological scaffold, and The feature is that the blood transport system of the blood vessel is simulated inside the biological stent, and the nutrients required for the tissue culture are transported to the inside of the biological stent by the simulated blood vessel.

Wherein, the simulated vascular system is a space that is mostly linearly extended inside the biological stent.

(10) ‧‧‧Multiple supply bioreactors

(20)‧‧‧Biological stent

(30) ‧‧‧First Supply Stream

(40) (40’) ‧‧‧Second Supply Stream

(31)‧‧‧External cavity parts

(32) ‧‧‧End pieces

(33) ‧‧‧Confined supply space

(34) ‧‧‧First inlet channel

(35) ‧‧‧First outflow channel

(321) ‧‧‧End caps

(322)‧‧‧Connectors

(41) (41’) ‧‧‧Second inlet channel

(42) (42’) ‧‧‧Second outflow channel

The first figure is a perspective view of a preferred embodiment of the present invention.

The second drawing is a partially exploded view of a preferred embodiment of the present invention.

Figure 3 is a cross-sectional view of a preferred embodiment of the present invention taken along the line sec of Figure 3-3.

Figure 4 is a cross-sectional view of a preferred embodiment of the present invention taken along the line 1-4 of Figure 4-4.

Figure 5 is a cross-sectional view of a preferred embodiment of the present invention taken along the line 5-1 of Figure 5-5.

Figure 6 is a schematic view of a perfusion flow according to a preferred embodiment of the present invention, showing a schematic diagram of fluid forming an external flow outside the bioscaffold. .

Figure 7 is a schematic view of a perfusion flow according to a preferred embodiment of the present invention, showing a schematic diagram of fluid forming an internal flow inside the bioscaffold.

Figure 8 is a schematic illustration of a perfusion flow in accordance with a preferred embodiment of the present invention showing a schematic representation of another internal flow of fluid within the bioscaffold.

In the following, a preferred embodiment of the invention will be further described with reference to the drawings.

First, referring to the first to fifth figures, a multi-supply flow bioreactor (10) is provided in a preferred embodiment of the present invention, which mainly comprises a biological support. (20), a first supply flow (30) and two second supply flow (40) (40').

The biological scaffold (20) is composed of an organic material such as glycolide lactide copolymer (Polyglactin 910), polyglycolic acid (PGA), polylactic acid (PLA), hydroxyapatite or a columnar body made of a biodegradable material such as an inorganic material such as magnesium strontium calcium or the like, and a base for the growth of cells during tissue engineering, and a large number of internal structures are formed therein. The three-dimensional space enables the cells to exchange gas, absorb nutrients and excrete metabolites, but the specific technical content or the materials used therein are not the technical features of the present invention, and are known technologies of the prior art. Therefore, in this embodiment, it is no longer redundant.

The first supply flow portion (30) has a hollow tubular outer cavity member (31) of a suitable length and inner diameter, and the biological support (20) is accommodated in an inner tube space, and the two end members (32) are respectively disposed on The ends of the tube shaft of the outer chamber member (31) are such that the nozzles at both ends of the tube member of the outer chamber member (31) are closed by the end members (32) to form a space inside the tube to be isolated from the outside. And surrounding a closed supply flow space (33) on the circumference of the biological support (20), and the two first inlet flow paths (34) are respectively disposed on the wall of one end of the tube shaft of the outer cavity member (31) to communicate Inside and outside the supply space (33), two first outflow channels (35) are respectively disposed on the wall of the outer end of the outer tube member (31) and communicate with the supply flow space (33). According to this, the external fluid can enter the supply space (33) via the first inlet passages (34), and then flow out of the outer chamber member (31) from the first outlet passages (35), thereby Forming an external supply flow flowing outside the biological support (20); further, each of the end pieces (32) has an end cover (321) respectively disposed at an axial end of the outer cavity member (31) On the nozzle, a columnar joint (322) is coaxially accommodated in The outer chamber member (31) and interposed between the corresponding end cap with the biological support (20) (321).

The supply flow is formed outside the biological support (20) with respect to the first supply flow (30), and each of the second supply flow (40) (40') is used for the biological support (20) The inside forms a supply flow, so that the inside and the outside of the biological support (20) can be respectively placed in an environment of different conditions, so that the tissue engineering is easy to cultivate, specifically: the second supply flow (40) A second inlet channel (41) and a second outlet channel (42) are disposed on each of the end members (32) and pass through an internal space on the column axis of the columnar biological support (20). Communicating with each other, so that the external fluid enters the inner space of the biological support (20) via the second inlet channel (41), and then flows out of the outer cavity member (31) via the second outlet channel (42). Thereby forming a supply flow flowing inside the biological support (20); the second supply flow (40') has a second inlet channel (41') and a second outlet channel (42'), The system is disposed on each of the end pieces (32), and communicates with each other via a plurality of axially extending and annularly arranged spaces inside the columnar biological support (20), thereby making the exterior After entering the majority of the space inside the biological support (20) via the second inflow channel (41'), the body flows out of the outer cavity member (31) through the second outflow channel (42'). Thereby a further supply stream that flows inside the bioscaffold (20) is formed.

By using the composition of the above components, the multi-feedstream bioreactor (10) is used as shown in the sixth to eighth figures, taking the tissue culture of the bone as an example, and is located in the supply space. The biological scaffold (20) in (33) can be externally formed on the peripheral side of the biological scaffold (20) by using the first supply flow portion (30) as shown in FIG. The flow provides the nutrients required for the growth of the skeletal cells, and regulates the conditions thereof, and can be formed inside the biological support (20) by using the second supply flow portion (40) as shown in the seventh figure. The supply stream provides the nutrients needed for the growth of bone marrow cells while regulating their conditions.

However, in the cultivation of large tissues such as bones, in view of the limitations of the prior art on the supply of cell nutrients, the present invention is effective in reinforcing the deficiencies of the prior art, which is specifically as shown in the eighth figure. As shown, another internal supply flow formed by the second supply flow portion (40') in a plurality of linearly extending spaces inside the biological stent (20) serves as a supply of vascular cell implantation and growth nutrients thereof, A simulated blood transport system is formed inside the biological scaffold (20) to provide a passage for nutrient transport, so that large-volume tissue culture can be smoothly performed, in other words, the second supply flow portion (40) and the first The second supply flow portion (40') can be used as a flow channel for different solutions in actual use, so that the multi-feed flow bioreactor (10) is used, through the first supply flow portion (30) Each of the second supply flow portions (40) (40') causes a solution of three different constituent components to flow, so that nutrients required for different cell growth can be simultaneously present in different portions of the biological scaffold (20), thereby facilitating application of the Biological stent (20) for different fine The culture, the culture of bone and other tissue to be carried out smoothly.

(20)‧‧‧Biological stent

(30) ‧‧‧First Supply Stream

(40) (40’) ‧‧‧Second Supply Stream

(31)‧‧‧External cavity parts

(32) ‧‧‧End pieces

(33) ‧‧‧Confined supply space

(34) ‧‧‧First inlet channel

(35) ‧‧‧First outflow channel

(321) ‧‧‧End caps

(322)‧‧‧Connectors

(41) (41’) ‧‧‧Second inlet channel

(42) (42’) ‧‧‧Second outflow channel

Claims (12)

A multi-supply flow bioreactor comprises: a biological support; a first supply flow portion having a supply flow space surrounding the circumference of the biological support, a first inlet channel and a The first outflow channel is respectively connected to the supply flow space, so that the supply flow space connects the first inlet flow channel and the first outlet flow channel; and a second supply flow portion has a second inlet flow channel One end is in communication with a predetermined space inside the biological stent, and a second outlet passage is connected at one end to a predetermined space inside the biological stent, and the second inlet channel is connected to the predetermined space inside the biological stent. The second outflow channel. The multi-supply flow bioreactor according to claim 1, wherein the first supply flow portion has a hollow outer cavity member for accommodating the biological support inside the hollow. The multi-feedstream bioreactor according to claim 2, wherein the first inflow channel and the first outflow channel are respectively disposed on the outer cavity member. A multi-feedstream bioreactor according to the scope of claim 2, wherein the outer chamber member is tubular. The multi-supply flow bioreactor according to claim 4, wherein the first supply flow system further comprises two end pieces, which are respectively disposed at two ends of the outer cavity tube shaft for closing The space inside the tube of the outer cavity member is isolated from the outside, thereby forming the supply space. a multi-feedstream bioreactor according to claim 5, wherein each of the The end piece has an end cover respectively, and the cover is disposed at an axially corresponding end of the outer cavity member, so that the two ends of the outer tube member are closed by the end caps, and a joint is received outside the outer cover member. In the cavity member, between the biological support and the corresponding end cover. The multi-feedstream bioreactor according to claim 6, wherein the second inflow channel and the second outflow channel are respectively disposed on each of the end pieces. The multi-feedstream bioreactor according to claim 1, wherein the number of the second supply stream is two. According to the multi-supply flow bioreactor according to claim 8, wherein each of the second supply flow portions respectively communicates with the second inflow channel and the corresponding portion by different spaces inside the biological support Two out of the runner. The multi-feedstream bioreactor according to claim 1, wherein the second inflow channel and the second outflow channel are respectively connected to a plurality of different spaces inside the bio-scaffold. A tissue culture method is a tissue culture using a biological scaffold, and is characterized in that a blood transport system of a blood vessel is simulated inside the biological scaffold, and the nutrient required for growth is transmitted to the simulated blood vessel to the The interior of the biological scaffold. The method of tissue culture according to claim 11, wherein the simulated vascular system is a space extending most linearly inside the biological stent.