JP2023124920A - hollow fiber bioreactor - Google Patents

Abstract

To provide a method of conveying a hollow fiber bioreactor used by implanting in a body and a liquid factor to an ischemic limb for sufficiently reacting a liquid factor to an ischemic limb.SOLUTION: A device is used by being implanted in a body, and has bundles of hollow fibers having a lumen capable of housing cells and a main body portion that stores the bundle of hollow fibers, wherein each hollow fiber is constituted such that does not release cells injected into the lumen but is composed of a semi-permeable membrane so as to permeate the liquid factor that the cells produce, and at least a part of the main body is constituted of a porous material so as to permeate the liquid factor that the cells injected in the lumen produce, but both ends of the hollow fiber bundles are fixed liquid tightly with the main body, one end and the other end of the bundle of the hollow fibers are connected with two tube bodies so such that each lumen is liquid densely communicated, the other ends of the two tube bodies are communicated with a liquid driver arranged outside of the body, the hollow fiber lumen contains cultured liquid including cells, due to the liquid driver, the cultured liquid circulates in the lumen of the hollow fibers.SELECTED DRAWING: Figure 3

Description

The present invention relates to hollow fiber bioreactors and methods for delivering liquid factors.

In recent years, attempts have been made to transplant various cells to repair damaged tissues and the like. An example of repairing damaged tissue by cell transplantation is CD34-positive cell therapy for severe lower limb diseases. In such treatment, CD34-positive cells are dissolved in physiological saline and injected intramuscularly into dozens of locations in both legs that are in a severe ischemic state (for example, see Non-Patent Document 1).
Bioreactors are used for cell culture and come in various types, including hollow fiber bioreactors. The lumen (inside) of the hollow fiber is called the IC, and the outside of the hollow fiber is called the EC, and the liquid (culture solution) is circulated through each separately.

As a bioreactor, a tissue cell culture system is described that performs efficient early proliferation of cell tissue in the injured area, that is, in vivo, and avoids the occurrence of bacterial infection in the injured area during treatment. (For example, Patent Document 1). Further, a hollow fiber bundle module is described in which a plurality of hollow fibers are arranged in parallel in a state in which the flexibility of the hollow fibers is suppressed by a fixing material so that the fibers can be maintained in a substantially straight line (for example, Patent Document 2). . Furthermore, a bioreactor has been described in which a culture solution is supplied through a medium supply membrane and cells are grown on the medium supply membrane and in a space outside the medium supply membrane (for example, Patent Document 3).

Patent No. 4067419 Japanese Patent Application Publication No. 2001-137669 Special Publication No. 6-102013

Fujita et al., Circulation Journal Vol.78, February 2014: 490-501

An object of the present invention is to provide a hollow fiber bioreactor that is used by being implanted in the body, and a method for delivering a liquid factor to an ischemic limb, in order to have a sufficient effect of a humoral factor on an ischemic limb. .

As a result of extensive studies, the present inventor has found that the above objective can be achieved by adopting a bioreactor having a hollow fiber and a main body that can house cells and allow humoral factors released from the cells to pass through. They discovered this and completed the present invention.

That is, the present invention relates to the following.
(1) A device that is used by being implanted in the body, and has a bundle of hollow fibers having a lumen that can store cells and a main body that stores the bundle of hollow fibers,
Each hollow fiber is composed of a semipermeable membrane that does not release cells injected into the lumen, but allows humoral factors produced by the cells to pass through.
At least a portion of the main body is made of a porous material so as to be permeable to humoral factors produced by cells injected into the lumen;
Both ends of the hollow fiber bundle are fixed to the main body in a liquid-tight manner, and one end and the other end of the hollow fiber bundle are connected to two tubular bodies so that their inner cavities communicate with each other in a liquid-tight manner. The other end of the tubular body is connected to a liquid drive unit placed outside the body,
A device in which the hollow fiber lumen contains a culture solution containing cells, and the culture solution is circulated through the hollow fiber lumen by a liquid drive unit.
(2) The device according to (1), wherein the main body is a porous membrane case.
(3) The device according to (1), wherein each hollow fiber is covered with a porous material.
(4) The device according to (3), wherein the porous material is expanded polytetrafluoroethylene (ePTFE).
(5) The two tubular bodies inside the body are connected to the liquid drive unit outside the body via a port-like structure fixed to the outside surface of the body, according to any one of (1) to (4). device.
(6) A method of delivering humoral factors to an ischemic limb, the method comprising:
providing a device to be implanted in the body, wherein the device to be implanted in the body includes a bundle of hollow fibers having a lumen capable of accommodating cells and a body portion housing the bundle of hollow fibers; Each hollow fiber is composed of a semi-permeable membrane so as not to release the cells injected into the lumen but to allow humoral factors produced by the cells to pass therethrough, and at least a portion of the main body. is made of a porous material that allows humoral factors produced by cells injected into the lumen to pass through, and both ends of the bundle of hollow fibers are fixed fluid-tightly to the main body.
filling the hollow fiber lumen with cells;
connecting both ends of the bundle of hollow fibers, the main body, and one tubular body in a liquid-tight manner;
storing the main body connected to the tubular body in a sheath that can be punctured into the body;
puncturing and indwelling the main body housed in the sheath near the lower limb artery through the incision on the body surface;
removing the sheath from the body portion by peeling the sheath through another incision on the body surface;
connecting the main body and another tubular body in a liquid-tight manner;
connecting the two tubular bodies connected to the main body with a liquid drive unit disposed outside the body, and
The method comprises the step of sending the culture medium into the hollow fiber lumen by means of a liquid drive.
(7) The method according to (6), further comprising the step of circulating the culture medium through the hollow fiber lumen.

According to the present invention, a bioreactor having a hollow fiber and a main body that can house cells and allow humoral factors released from the cells to pass through is employed, and is implanted near the outside of an artery in the lower limb, thereby allowing cells to be absorbed. The released humoral factors can be sufficiently applied to the target ischemic limb.
In particular, by inserting the device body into the sheath and then inserting the device by puncturing the device through the opening, it is relatively easy to insert the device into the vicinity of the outside of the artery of the lower limb.

FIG. 1 is an overview of a device according to a preferred embodiment of the invention. (a) is an overall view of a device according to a preferred embodiment of the invention, the body being connected to a liquid drive via a port; (b) is an overall view of a device according to a preferred embodiment of the invention, with the body separated from the liquid drive at the port; FIG. 2 shows one embodiment of the hollow fiber housed in the main body of the device of the present invention. FIG. 3 is an overview of a device according to a preferred embodiment of the invention, further comprising a fresh liquid supply and a waste liquid storage. FIG. 4 is a schematic diagram illustrating a preferred embodiment of the treatment method of the present invention. (a) shows the step of storing the device connected to the tubular body in a sheath that can be inserted into the body. (b) shows the step of making two incisions on the body surface. (c) shows the step of puncturing the device housed in the sheath near the lower limb artery from one incision toward the other. (d) shows removing the sheath from the implanted device by peeling the sheath from the other incision after the body of the device is in place. (e) shows the step of connecting two tubular bodies connected to the device with ports placed on the body surface.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the drawings.
First, the device of the present invention will be explained.

FIG. 1 is an overall view of a device according to a preferred embodiment of the invention. (a) shows the state after connection with the liquid drive unit 5, and (b) shows the state before connection with the liquid drive unit 5. In each figure, for convenience of explanation, in the description of the device 1, the side in the insertion direction near the outside of the artery is called the distal side, and the opposite side is called the proximal side.
As shown in FIG. 1, the device 1 includes a main body 2, a tubular body 3, a port 4, and a liquid driver 5. In the device 1, by placing the main body part 2 containing a cell composition containing cells near the outside of the artery of the lower limb, humoral factors are generated in the cells and released from the cells, and the humoral factors are delivered to the ischemic limb. It can improve blood flow in peripheral artery disease caused by arteriosclerosis, foot lesions caused by diabetes, and lower limb ischemia.

In the present invention, the term "cell" includes a "cell suspension" in which cells are suspended in any medium. The cells may be in the form of a single cell, or the cells may be connected to each other via an intervening substance. A cell may contain any gene or a viral vector containing any gene, or the like. Moreover, the cell may be a cell culture. In the present invention, the cell culture refers to what is obtained through a cell culture step, and includes, but is not limited to, spheroids, sheet-shaped cell cultures, and fragments of sheet-shaped cell cultures.
In the present invention, media for suspending cells include, but are not limited to, water, physiological saline, media, buffers, diluents, cell preservation solutions, and swelling bodies such as gels.

In the present invention, the location where the device is placed includes, for example, a site where an injury (wound) exists in a living body or the vicinity thereof. Damage (wound) includes, but is not limited to, damage to blood vessels, stenosis, and in such cases, the target site in the tissue is in the tissue adjacent to the blood vessel, preferably in the wall of the blood vessel. and the surrounding organizations. In one embodiment, the site of interest in the tissue is a vascular sheath around the vascular tissue. The target site in the tissue may be determined in advance by ultrasonic examination, CT examination, etc. before administration.
In one embodiment, the subject in whom the device of the invention is placed has a lower extremity disease. Diseases of the lower extremities include any diseases that affect the lower extremities. Examples of such diseases include, but are not limited to, peripheral artery disease, varicose veins, deep vein thrombosis, and diabetic foot lesions. Preferably, the lower extremity disease in the present invention is a disease in which blood vessels in the lower extremities are narrowed or occluded, resulting in worsening blood flow in the lower extremities, such as peripheral artery disease. In the present invention, peripheral arterial diseases include, but are not limited to, lower extremity arteriosclerosis obliterans, Buerger's disease, collagen disease, etc., and in particular severe lower extremity arteriosclerosis obliterans are treated as severe lower extremity ischemia. It is called.
As used herein, "near the outer side of arteries in the lower limbs" includes, for example, tissues around blood vessels in the lower limbs (e.g., anterior tibial artery, posterior tibial artery, or peroneal artery), vascular sheaths (thin membrane tissue on the surface of blood vessels), etc. refers to the location outside the blood vessels of the lower extremities.

Furthermore, in the present invention, the cells to be placed are not particularly limited, and include, for example, adherent cells (adherent cells). Adherent cells include, for example, adhesive somatic cells. Examples of somatic cells include myoblasts (e.g., skeletal myoblasts, etc.), muscle satellite cells, mesenchymal stem cells (e.g., bone marrow, adipose tissue, peripheral blood, skin, hair roots, muscle tissue, intrauterine cells, etc.). membranes, placenta, umbilical cord blood-derived, etc.), tissue stem cells such as cardiomyocytes, fibroblasts, and cardiac stem cells, embryonic stem cells, pluripotent stem cells such as iPS (induced pluripotent stem) cells, synovial cells, and chondrocytes. , epithelial cells (e.g., oral mucosal epithelial cells, retinal pigment epithelial cells, nasal mucosal epithelial cells, etc.), endothelial cells (e.g., vascular endothelial cells, etc.), hepatocytes (e.g., hepatic parenchymal cells, etc.), pancreatic cells (e.g., pancreatic islet cells, etc.), renal cells, adrenal gland cells, periodontal ligament cells, gingival cells, periosteal cells, skin cells, etc. Somatic cells may be those differentiated from iPS cells (iPS cell-derived cells), such as iPS cell-derived cardiomyocytes, fibroblasts, myoblasts, epithelial cells, endothelial cells, hepatocytes, pancreatic cells, and kidney cells. Examples include cells, adrenal gland cells, periodontal ligament cells, gingival cells, periosteal cells, skin cells, synovial cells, and chondrocytes. Cells include floating cells. Examples of floating cells include T lymphocytes and B lymphocytes.
In one embodiment, the cells of the invention are particularly preferably cells that can promote angiogenesis, eg, cells that can secrete factors that promote angiogenesis, eg, cytokines such as VEGF.

These cells may be derived from any mammal or other organism, such as, but not limited to, humans, non-human primates, dogs, cats, pigs, horses, goats, sheep, etc., for example. These cells can be found in separate states, in clusters of cells (spheroids), or in membranous state as sheet-like cell cultures, or in membranous sheet-like cell cultures. It is prepared by floating or suspending it in a liquid in the form of a crushed substance. A liquid containing cells provided in such a state is called a cell composition, and may contain other substances as long as they are applicable to the present invention. The liquid that can be used for floating or suspending cells is not particularly limited, but it is preferable that it does not harm living organisms. For example, various buffer solutions, culture solutions, etc. may be used alone or in combination of two or more. be able to. The cell composition may contain drugs such as therapeutic agents and anesthetics as appropriate. In this specification, when the term "cell" is used below, it may mean a cell suspension or a cell composition.

In addition, examples of the humoral factors released by the above-mentioned cells include cell growth factors, differentiation-inducing factors, hematopoietic factors, and other various cytokines. Examples of these humoral factors include HGF, VEGF, and SDF-1. Note that it is naturally possible that humoral factors other than the exemplified humoral factors are released from the above-mentioned cells, and since these released humoral factors act on the tissue as a whole, the term "humoral" in this specification refers to It goes without saying that the concept includes humoral factors other than those exemplified.

As shown in FIG. 1, the main body 2 has a tubular overall shape, and stores a bundle 25 of hollow fibers therein. In order to increase cell culture efficiency, it is preferable to maximize the number of hollow fibers 21 included in the hollow fiber bundle 25 and the cell culture surface area of the hollow fiber lumen, and to reduce invasion into the body, Preferably, the volume of the hollow fiber bundle 25 is minimized. For example, the hollow fiber bundle 25 housed in the main body 2 consists of approximately 10,000 to 12,000 hollow fibers, and the entire bundle of hollow fibers has a cell culture surface area of 1.5 to 3.0 m ² . can have. As a result, a cell culture density of approximately 200 million cells per mL can be maintained.

As shown in FIG. 2, each hollow fiber 21 is constituted by a tubular membrane (hollow fiber membrane) having a lumen 22 formed to have an inner diameter through which a liquid can flow. Therefore, each hollow fiber 21 has cells in its lumen, and cells 300 can be cultured within the hollow fiber 21 by supplying a liquid medium from the liquid drive unit. The lumen of the hollow fiber is also referred to as the "IC space." Each hollow fiber is composed of a semi-permeable membrane that does not release cells injected into the lumen but allows humoral factors produced by the cells to pass through, and cells adhere to the inner wall of the hollow fiber. , or can multiply while floating.

The material of the hollow fiber is not limited as long as it is a semi-permeable membrane that does not release cells and is permeable to humoral factors such as cytokines, but examples include hydrophilic resins such as polyethersulfone and resin materials such as polypropylene. It is possible to use a resin material whose surface has been treated with hydrophilic treatment. Preferred hollow fiber materials include polysulfone (PS), polyethersulfone (PES), polyester polymer alloy (PEPA), ethylene vinyl alcohol copolymer (EVAL), polymethyl methacrylate (PMMA), and polyacrylonitrile copolymer ( PAN) can be used. The hollow fibers can also be coated with an outer hollow fiber porous membrane 24 of a porous material such as expanded polytetrafluoroethylene (ePTFE).

The hollow fibers 21 can have, for example, an outer diameter of 250 μm, a wall thickness of 10 μm to 50 μm, and an inner diameter of 200 μm. Cells housed in the inner diameter of the hollow fiber can be in a single cell state, a clump-like state (spheroid), or a membranous state, as long as they can grow by adhering to or floating on the inner diameter of the hollow fiber. It may be in any state of crushed sheet-shaped cell culture.

Further, as shown in FIG. 1, the main body portion 2 is liquid-tightly fixed to the distal end 26 and the proximal end 27 of the hollow fiber bundle 25. The hollow fiber outer space 23 is a space formed inside the tubular main body 2 and outside the hollow fiber bundle 25 housed within the main body 2, and is also referred to as an "EC space." Further, the distal end 26 and proximal end 27 of the bundle of hollow fibers are also in liquid-tight communication with the tubular body 3. The tubular body 3 is in liquid-tight communication with the liquid driver 5 using a port 4 present on the external surface of the body.

By the way, at least a part of the main body part 2 that accommodates the bundle of hollow fibers is made of a member through which humoral factors released from cells can pass. Thereby, when the device 1 is placed near the outside of the artery, the cells housed in the hollow fiber lumen 22 release humoral factors, but the humoral factors are released outside the device 1. Then, the released humoral factors act on a target site in the artery of the lower limb, for example, an ischemic site. On the other hand, the cells are not released outside the hollow fiber, and are housed in the lumen 22 near the outside of the artery. This prevents cells from spreading near the outside of the artery and allows the humoral factor to sufficiently reach the target site.

The member constituting at least a part of the main body is not particularly limited as long as it allows the target humoral factor to pass through and does not allow cells to pass through. Materials that have pores that allow cells to pass through but do not allow cells to pass through, specifically fibrous materials (mesh fabrics) with a predetermined mesh such as woven fabrics (woven fabrics, knitted fabrics) and nonwoven fabrics, semipermeable membranes, etc. Porous membranes and the like can be used alone or in combination of two or more. In particular, porous membranes (porous PTFE membranes) made of fluororesin mainly composed of polytetrafluoroethylene (registered trademark) are chemically stable and have excellent heat resistance, chemical resistance, and mechanical strength. Therefore, it can be suitably used.

As described above, the main body 2 of the device 1 has the distal end 26 and the proximal end 27 of the hollow fiber bundle 25 sealed, and is made of a material that does not allow cells to pass through. Therefore, the device 1 does not release the stored cells to the outside. Therefore, even if the stored cells are left in the body cavity for a relatively long period of time, the cells are prevented from spreading outside the device 1, and the humoral factors are delivered to the desired part of the tissue such as the ischemic area. Delivery becomes more reliable. The distal end 26 and proximal end 27 of the hollow fiber bundle 25 can be liquid-tightly fixed to the main body by, for example, potting.

The liquid to be circulated (culture medium) is delivered from the liquid driver 5 through the port 4 and the tubular body 3 into the lumen of the hollow fiber (ie, the "IC space"). Thereafter, the culture medium, together with humoral factors produced by the cells present in the lumen of the hollow fiber, is further transferred to the outer space 23 of the hollow fiber (i.e., the "EC space"), and further to the blood vessels in the body outside the main body 2. It reaches the vicinity of the outside or passes through the tubular body 3 and port 4 to the liquid driver 5 as waste liquid. The culture solution sent from the liquid drive unit 5 may be added with nutrients for the cells to be cultured.
The number and surface area of the hollow fibers 21 included in the hollow fiber bundle 25 housed in the main body 2 are maximized, and the volume of the hollow fiber bundle 25 is minimized, so that the outer space 23 of the hollow fibers (i.e. The "EC space") is minimized, and most of the culture fluid sent from the liquid driver 5 is delivered to the "IC space." The length and width of the main body part 2 are not particularly limited as long as it can be placed near the outside of the lower limb, but may be, for example, 30 mm to 100 mm in length and 3 mm to 10 mm in width.

The tubular body 3 is a sterile tube configured to be able to deliver a cell culture solution, and has an appropriate length for connection from the device body to the port 4 outside the body.
The port 4 can be used on the external surface of the body to connect the tubular body 3 and the liquid drive 5, for example a port used for joining sterile tubes or using a sterile tube joining device. I can do it.
As the liquid drive unit 5, a device capable of circulating cell culture fluid, such as an infusion pump, can be used.

As shown in FIG. 3, the device 1 may have a new liquid supply section 6 and a waste liquid storage section 7 in addition to the liquid drive section 5 outside the body. Thereby, the culture medium can be rapidly supplied into the hollow fibers, and the culture medium after circulation can be stored as waste liquid.

In addition, it goes without saying that the size of each component of the device 1 described above can be appropriately set by those skilled in the art according to the condition of the blood vessels of the lower limbs to which it is applied, even if there is no particular mention.

Next, the treatment method of the present invention will be explained based on preferred embodiments.
FIG. 4 is a schematic diagram illustrating a preferred embodiment of the treatment method of the present invention. Note that in the related drawings, for the purpose of explaining the present embodiment, some parts may be enlarged or reduced and shown at a ratio different from the actual size.

The treatment method of the present invention is a method for delivering humoral factors to an ischemic limb, comprising:
providing a device to be implanted in the body, wherein the device to be implanted in the body includes a bundle of hollow fibers having a lumen capable of accommodating cells and a body portion housing the bundle of hollow fibers; Each hollow fiber is composed of a semi-permeable membrane so as not to release the cells injected into the lumen but to allow humoral factors produced by the cells to pass therethrough, and at least a portion of the main body. It is made of a porous material that allows humoral factors produced by cells injected into the lumen to pass through.
filling the hollow fiber lumen with cells (cell filling step);
a step of connecting both ends of the bundle of hollow fibers, the main body, and one tubular body in a liquid-tight manner (connection step);
storing the main body connected to the tubular body in a sheath that can be punctured into the body (sheath storage step);
a step of puncturing and indwelling the main body housed in the sheath near the lower limb artery through an incision on the body surface (indwelling step);
removing the sheath from the main body by peeling the sheath from another incision on the body surface (sheath peeling step);
a step of connecting the main body and another tubular body in a liquid-tight manner (tubular body connecting step);
a step of connecting the two tubular bodies connected to the main body with a liquid drive unit disposed outside the body (liquid drive unit connection step);
The method includes a step of sending a culture solution to the hollow fiber lumen by a liquid drive unit (culture solution delivery step).

Hereinafter, in this embodiment, the device 1 described above is placed near the outside of the lower limb artery 100 200 to treat an ischemic limb.

First, in a first step, a device 1 having a main body 2 is provided. As described above, the main body part 2 houses a hollow fiber and a bundle of hollow fibers having a lumen capable of accommodating cells, and each hollow fiber does not release the cells injected into the lumen, but the cells It is composed of a semi-permeable membrane that allows the produced humoral factors to pass through. Further, the main body portion 2 is made of a member through which humoral factors released from cells can pass. The main body portion 2 may be appropriately cleaned and sterilized using known methods.

Next, in the cell filling step, cells are introduced into the lumen of each hollow fiber. The tips or ends of the bundle of hollow fibers are open before being filled with the cell composition, but after being filled with the cell composition, they are sealed and housed in the main body 2, so that they are spatially connected to the outside of the hollow fibers. It is configured so that it does not lead to

Next, in the connecting step, both ends of the hollow fiber bundle 25, the main body portion 2, and the tubular body 3 are connected so as to fluid-tightly communicate with each other. For example, the main body portion 2 is fluid-tightly fixed to the distal end 26 and proximal end 27 of the hollow fiber bundle 25 by potting in which resin is injected and hardened. The tubular body 3 is connected to the main body 2 via, for example, a connector so as to communicate liquid-tightly.

Next, in the sheath storage step, the main body portion connected to the tubular body is stored in a sheath 400 that can be inserted into the body. As shown in FIG. 4, the sheath 400 is hard enough to be inserted into the body, and has a sharp tip. Since the sheath 400 is configured to be able to be inserted into the body, the main body portion may be stored in close contact with the sheath 400. Preferred sheath materials include polyolefins such as polyethylene, polypropylene, polybutene, ethylene-propylene copolymers, ethylene-vinyl acetate copolymers, ionomers, or mixtures of two or more of these, flexible polyvinyl chloride resins, polyamides, and polyamides. Examples include elastomers, polyesters, polyester elastomers, polyurethanes, fluororesins such as polytetrafluoroethylene, silicone rubbers, latex rubbers, and the like.

Next, in the indwelling step, as shown in FIG. 4, the main body 2 housed in the sheath is punctured near the lower limb artery through the incision 500 on the body surface and indwelled. The incision 500 and the space 600 for placing the device can be formed by normal surgical techniques used in lower extremity incisions to the extent necessary to puncture the main body housed in the sheath. I can do it.
In addition, in order to have enough strength to withstand the puncture of the main body part 2 housed in the sheath, a hard material is used so that both ends of the bundle of hollow fibers housed in the main body part 2 and the part between them can also be punctured. It may be fixed to provide stability.
In this step, the device 1 arranged as described above is left in place for a certain period of time, for example, 2 to 60 days, preferably 7 to 21 days.
In this step, by indwelling the device 1, liquid factors produced and released in the cells 300 are released near the outside of the blood vessel via the main body 2. The released fluid factors are expected to act on the ischemic tissue and treat the ischemic limb.

Next, in the sheath peeling step, as shown in FIG. 4, the sheath 400 is exposed to the body surface through an opening 500 different from the punctured opening 500, and the sheath is peeled and removed from the main body. The sheath has a peel line formed in advance in the same direction as the direction of puncture into the body (that is, in the longitudinal direction), and can be divided into a plurality of pieces in the longitudinal direction and removed from the main body.

Next, in the tubular body connecting step, the main body portion 2 and another tubular body 3 are connected so as to communicate in a liquid-tight manner. The tubular body 3 is connected to the main body 2 via, for example, a connector so as to communicate liquid-tightly.

Next, in the liquid drive unit connecting step, the two tubular bodies 3 connected to the main body 2 are connected to the liquid drive unit 5 disposed outside the body. The connection between the tubular body and the liquid drive can be made via a port 4 on the external surface of the body. For example, a port used for joining sterile tubes or a sterile tube joining device can be used. As the liquid drive unit 5, a device capable of circulating cell culture fluid, such as an infusion pump, can be used.

Next, in the culture solution delivery step, the culture solution is delivered to the hollow fiber lumen by the liquid drive unit 5. The culture medium can contain nutrients for the cells.
In addition, if necessary, the step of circulating a culture solution into the hollow fiber lumen for cell culture may further be included, and new cells are injected from outside the body, circulated, and supplied into the hollow fiber. It can be configured as follows.

Next, in the removal step, the device 1 is removed from near the outside of the artery.
In this step, first, an incision is made again as appropriate to expose the tubular body 3 connected to the main body part 2.
Next, the main body 2 is pulled out along with the tubular body 3, and the device 1 is removed from near the outside of the artery.
Finally, suture the incision.

As described above, according to the present invention, cells can be easily indwelled in a body cavity by employing a device having a main body that can house cells and allow humoral factors released from the cells to pass through. can. As a result, the liquid factors released from the indwelled cells can be made to sufficiently act on the target tissue.
In particular, by employing a tubular device main body, it is relatively easy to insert the cell retention device into the vicinity of the outside of the artery.

In particular, according to the invention, not only cells used in normal tissue grafts or cell transplants, but also cells (or grafts containing cells) which are not suitable for transplantation but release the necessary fluid factors, can be used. Available for use.

Although the present invention has been described above with reference to the illustrated embodiments, the present invention is not limited thereto.
In the present invention, each configuration can be replaced with any one that can perform the same function, or any configuration can be added.

1 Device 2 Main body (porous membrane case)
21 Hollow fiber 22 Hollow fiber lumen (inner diameter)
23 Space outside the hollow fiber 24 Porous membrane outside the hollow fiber 25 Bundle of hollow fibers 26 Tip of the bundle of hollow fibers 27 Base end of the bundle of hollow fibers 3 Tubular body 4 Port 5 Liquid drive unit 6 New liquid supply unit 7 Waste liquid storage Part 100 Artery 200 Near the outside of the artery 300 Cell 400 Sheath 500 Incision 600 Space (inside the body)

Claims (7)

A device used by being implanted in the body, having a bundle of hollow fibers having a lumen capable of housing cells, and a main body portion housing the bundle of hollow fibers,
Each hollow fiber is composed of a semipermeable membrane that does not release cells injected into the lumen, but allows humoral factors produced by the cells to pass through.
At least a portion of the main body is made of a porous material so as to be permeable to humoral factors produced by cells injected into the lumen;
Both ends of the hollow fiber bundle are fixed to the main body in a liquid-tight manner, and one end and the other end of the hollow fiber bundle are connected to two tubular bodies so that their inner cavities communicate with each other in a liquid-tight manner. The other end of the tubular body is connected to a liquid drive unit placed outside the body,
A device in which the hollow fiber lumen contains a culture solution containing cells, and the culture solution is circulated through the hollow fiber lumen by a liquid drive unit. 2. The device of claim 1, wherein the body is a porous membrane case. 2. The device of claim 1, wherein each hollow fiber is covered with a porous material. 4. The device of claim 3, wherein the porous material is expanded polytetrafluoroethylene (ePTFE). The device according to any one of claims 1 to 4, wherein the two tubular bodies inside the body are connected to a liquid drive outside the body via a port-like structure fixed to the external surface of the body. A method of delivering humoral factors to an ischemic limb, the method comprising:
providing a device to be implanted in the body, wherein the device to be implanted in the body includes a bundle of hollow fibers having a lumen capable of accommodating cells and a body portion housing the bundle of hollow fibers; Each hollow fiber is composed of a semi-permeable membrane so as not to release the cells injected into the lumen but to allow humoral factors produced by the cells to pass therethrough, and at least a portion of the main body. is made of a porous material that allows humoral factors produced by cells injected into the lumen to pass through, and both ends of the bundle of hollow fibers are fixed fluid-tightly to the main body.
filling the hollow fiber lumen with cells;
connecting both ends of the bundle of hollow fibers, the main body, and one tubular body in a liquid-tight manner;
storing the main body connected to the tubular body in a sheath that can be punctured into the body;
puncturing and indwelling the main body housed in the sheath near the lower limb artery through the incision on the body surface;
removing the sheath from the body portion by peeling the sheath through another incision on the body surface;
connecting the main body and another tubular body in a liquid-tight manner;
connecting the two tubular bodies connected to the main body with a liquid drive unit disposed outside the body, and
The method comprises the step of sending the culture medium into the hollow fiber lumen by means of a liquid drive. 7. The method of claim 6, further comprising circulating culture fluid through the hollow fiber lumen.