TWM517199U - Cell carrying device and cell culture system - Google Patents

Description

Cell carrying device and cell culture system

The present invention relates to a cell carrying device, and more particularly to a cell carrying device comprising a polyurethane elastic film, a supporting structure and at least one channel, and a cell culture system comprising the cell carrying device.

Since many cells in an organism grow in a dynamic environment, for example, the walls of the heart, lungs, or stomach expand and contract with pressure, and the muscle or skin surface stretches with pressure, so when When the cells are observed in vitro, it is necessary to simulate a dynamic environment in accordance with the organism to carry and culture the cells.

Existing devices that provide a dynamic environment for observing cells use a resilient silicone film and apply an external force through a mechanical or fluid fluid to impart dynamic stimulation to cells carried on the silicone film. However, under long-term use, the silicone film tends to reduce the durability of the device due to the fatigue fatigue.

In addition, the device for observing adherent cells not only needs to provide a good cell growth environment, but also needs to have a good cell adhesion effect. Therefore, it is usually necessary to additionally perform chemical modification or protein coating to enhance cell adhesion.

Therefore, it is an object of the present invention to provide a cell-carrying device suitable for observing adherent cells.

Therefore, the novel cell carrying device comprises an elastic film, a supporting structure and at least one channel, wherein the elastic film is made of polyurethane. The elastic membrane is used to carry cells. The support structure is fixed to the periphery of the elastic film. The at least one channel is connected to the bottom of the elastic film to apply stress to the elastic film.

Therefore, another object of the present invention is to provide a cell culture system comprising the cell carrying device as described above, and an adherent cell attached to the elastic film.

The effect of the novel is that the polyurethane elastic membrane of the cell carrying device has good elasticity and durability, and can improve cell adhesion without additional chemical modification or protein coating.

The present invention will be described in detail below. Preferably, the support structure is fixed to opposite sides of the periphery of the elastic film.

In a first embodiment of the present invention, the cell carrying device further includes a base spaced apart from the bottom of the elastic film, and the at least one hole is penetrating the base.

In a second embodiment of the present invention, the cell carrying device further includes a carrier column disposed at a bottom of the elastic film and spaced apart from the support structure, and the elastic film is partially attached to the carrier column.

Preferably, the polyurethane is such that the polyether polyol and the aliphatic group are The isocyanate reaction is followed by reaction with a chain extender. More preferably, the polyether polyol is selected from the group consisting of polyethylene glycol (PEG), polypropylene glycol (PPG), or a combination thereof. More preferably, the molar ratio of PEG to PPG is 3:7 or more. The aliphatic polyisocyanate avoids the risk of toxicity that aromatic compounds may have. More preferably, the aliphatic polyisocyanate is selected from the group consisting of isophorone diisocyanate (IPDI), hexamethylene diisocyanate (HDI), or a combination thereof. The chain extender is selected from the group consisting of 1,4-butanediol (1,4-BD), ethylene glycol, or a combination thereof.

More preferably, the molar ratio of the aliphatic polyisocyanate, the polyether polyol and the chain extender ranges from 2:1:1 to 3:2:1 to obtain good elasticity (high elongation) and durability ( High strength polyurethane).

According to the cell culture system of the present invention, the adherent cells include those which are readily available to those skilled in the art (for example, those available from domestic or foreign hosting institutions), or utilize the art. An adherent cell strain isolated and purified from a natural source by a conventional cell separation method. Preferably, the adherent cells are selected from the group consisting of mesenchymal stem cells, fibroblasts, epithelial cells, endothelial cells, Astrocytic cells, kidney cells, hepatocytes, epidermal cells, corneal cells, and combinations thereof. More preferably, the adherent cells are selected from the group consisting of placenta-derived mesenchymal stem cells (eg, human placenta-derived mesenchymal stem cells), embryonic kidney cells (eg, human embryonic kidney cells). Cell line HEK293), embryonic fibroblast (eg, mouse embryonic fibroblast cell line 3T3), or a combination thereof.

1‧‧‧cell bearing device

2‧‧‧elastic film

3‧‧‧Support structure

4‧‧‧ Holes

5‧‧‧Base

6‧‧‧Loading column

9‧‧‧ Attached cells

Other features and effects of the present invention will be apparent from the following description of the drawings. FIG. 1 is a cross-sectional view showing the structure of the cell carrying device of the first embodiment of the present invention; FIG. 2 is a cross-sectional view. The structure of the cell carrying device of the first embodiment under the application of the vibration stress in the vertical direction; FIG. 3 is a cross-sectional view showing the structure of the cell carrying device of the second embodiment of the present invention; and FIG. 4 is a cross-sectional view illustrating the The cell carrying device of the second embodiment is configured to apply a tensile stress in the horizontal direction.

Before the present invention is described in detail, it should be noted that in the following description, similar elements are denoted by the same reference numerals.

The present invention will be further described in the following examples, but it should be understood that these examples are for illustrative purposes only and are not to be construed as limiting.

<Preparation of elastic film>

IPDI, PEG1000 (weight average molecular weight of 1,000) and PPG1000 (weight average molecular weight of 1,000) were mixed at a molar ratio of 2:0.5:0.5, and reacted at 70 ° C for 2 hours to obtain a prepolymer, and then added. 1,4-BD (as a chain extender with a molar ratio of 1:2 to IPDI), reacted at 110 ° C for 24 hours, then dissolved in dimethyl acetamide (DMAc at a solids content of 20%) And coated on a release paper, finally dried to remove DMAc and tear off the release paper to obtain a polyurethane elastic film having a thickness of about 0.025 mm.

<Test of elastic film>

The polyurethane elastic film obtained above was subjected to an elongation test by ASTM D5034 and found to be 800%, showing that the polyurethane elastic film obtained above had a relatively high elongation.

The above-prepared polyurethane elastic membrane was used to culture human placenta-derived mesenchymal stem cells (experimental group), and the cultured culture dish was used as a control group, and cultured in an environment of 37 ° C, 5% CO ₂ for 7 days, and then the experiment was carried out. The viable cell densities measured in the group and the control group were 8.48×10 ⁴ /cm ² and 5.76×10 ⁴ /cm ² , respectively. The viable cell density of the experimental group was significantly increased by 47% compared with the control group. It is shown that the polyurethane elastic film obtained as described above can effectively promote the growth of adherent cells.

<Cell bearing device and cell culture system>

Referring to FIG. 1 and FIG. 2, the first embodiment of the novel cell carrying device 1 comprises an elastic film 2, a supporting structure 3, four holes 4 and a base 5. The elastic film 2 is a polyurethane elastic film obtained as described above, and the elastic film 2 is adhered with an adherent type cell 9 (the size is only a schematic), and can be used for emulating cells in a heart, lung or stomach that frequently expands and contracts. The dynamic growth environment of the wall.

The support structure 3 is two phases fixed to the periphery of the elastic film 2 The reverse side is such that the elastic film 2 is fixed and does not move.

The channels 4 are connected to the bottom of the elastic film 2, and are evacuated through the holes 4 by an external vacuum device (not shown) so that the elastic film 2 carries the portion of the adherent cells 9 The stress in the vertical direction is recessed downward; when the pumping is stopped, the elastic film 2 is returned to the non-recessed state before the pumping. By changing the pressure and frequency of the pumping, the elastic film 2 can vibrate between a recessed state and a non-recessed state, thereby providing the vertical type of vibration stress to the adherent cells 9 on the elastic film 2.

The base 5 is spaced apart from the bottom of the elastic film 2, and the holes 4 are penetrated through the base 5 to control the elastic film 2 to receive a uniform vertical direction stress during pumping.

Referring to FIG. 3 and FIG. 4, the second embodiment of the novel cell carrying device 1 comprises an elastic film 2, a supporting structure 3, a tunnel 4 and a supporting column 6. The elastic film 2 is a polyurethane elastic film obtained as described above, and the elastic film 2 is adhered with an adherent type cell 9 (the size is only a schematic) for simulating the dynamic growth environment of the muscle on the muscle or skin surface which is frequently stretched. .

The support structure 3 is fixed on opposite sides of the periphery of the elastic film 2 so that the elastic film 2 is fixed and does not move.

The hole 4 is connected to the bottom of the elastic film 2 and surrounds the carrier column 6, and is evacuated through the holes 4 by an external vacuum device (not shown), so that the elastic film 2 carries the adhesion type. The portion of the cell 9 is stretched outward by the stress in the horizontal direction; when the pumping is stopped, the elastic film 2 is returned to the non-stretched state before the pumping. By changing the pressure and frequency of the pumping, the elastic film 2 can be pulled between the stretched state and the non-stretched state. Stretching, thereby providing the tensile stress in the horizontal direction to the adherent cells 9 on the elastic film 2.

The supporting column 6 is disposed at the bottom of the elastic film 2, and is spaced apart from the supporting structure 3 through the hole 4, and the elastic film 2 is partially attached to the supporting column 6 to control the attached portion to be pumped. It is subjected to stress in the horizontal direction.

In summary, the novel cell-carrying device 1 can provide a good cell growth environment and cell adhesion effect by using the elastic and durable elastic film 2 without additional chemical modification or protein coating, and is suitable for observing adhesion. Type cell 9, so it can achieve the purpose of this new type.

However, the above is only the embodiment of the present invention, and when it is not possible to limit the scope of the present invention, all the simple equivalent changes and modifications according to the scope of the patent application and the contents of the patent specification are still This new patent covers the scope.

1‧‧‧cell bearing device

2‧‧‧elastic film

3‧‧‧Support structure

4‧‧‧ Holes

5‧‧‧Base

9‧‧‧ Attached cells

Claims (10)

A cell carrying device comprising: an elastic film for carrying cells; a support structure fixed to a periphery of the elastic film; and at least one channel communicating with the bottom of the elastic film to apply stress to the elastic film, wherein The material of the elastic film is polyurethane. The cell carrying device of claim 1, wherein the support structure is fixed to opposite sides of the periphery of the elastic film. The cell carrying device of claim 2, further comprising a base disposed at a bottom of the elastic film, and the at least one hole is penetrating the base. The cell carrying device of claim 2, further comprising a carrier column disposed at a bottom of the elastic film and spaced apart from the support structure, and the elastic film is partially attached to the carrier column. The cell carrying device according to claim 1, wherein the polyurethane is obtained by reacting a polyether polyol with an aliphatic polyisocyanate and then reacting with a chain extender. The cell-carrying device according to claim 5, wherein the aliphatic polyisocyanate is selected from the group consisting of isophorone diisocyanate, hexamethylene diisocyanate or a combination thereof. The cell-carrying device according to claim 5, wherein the chain extender is selected from the group consisting of 1,4-butanediol, ethylene glycol or a combination thereof. A cell culture system comprising the cell-carrying device according to claim 1, and an adherent cell attached to the elastic membrane. The cell culture system according to claim 8, wherein the adherent cell is selected from the group consisting of mesenchymal stem cells, fibroblasts, epithelial cells, endothelial cells, stellate cells, kidney cells, Hepatocytes, epidermal cells, corneal cells, and combinations thereof. The cell culture system according to claim 9, wherein the adherent cell is selected from the group consisting of placenta-derived mesenchymal stem cells, embryonic kidney cells, embryonic fibroblasts, or a combination thereof.