KR20150118279A - Polydimethylsiloxane-Based Apparatus for Three-dimensional Cell Culture - Google Patents
Polydimethylsiloxane-Based Apparatus for Three-dimensional Cell Culture Download PDFInfo
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Abstract
Description
본 발명은 폴리디메틸실록산 기반 3차원 세포 배양 장치에 관한 것으로, 구체적으로는 상부 표면에 3차원 구조를 가지는 배양부; 배양부와 하부 표면을 연결하는 모세관; 및 모세관을 채우는 충진용젤을 포함하는 3차원 배양 장치에 관한 것이다.
The present invention relates to a polydimethylsiloxane-based three-dimensional cell culture apparatus, and more particularly, to a culture apparatus having a three-dimensional structure on a top surface thereof; A capillary connecting the culture section and the lower surface; And a filling liquid filling the capillary tube.
최근 2차원으로 배양된 세포와 3차원으로 배양된 세포는 유전자 및 단백질 발현이 다르다는 연구결과들이 발표됨에 따라 3차원 세포배양기술에 대한 관심이 높아지고 있다. 세포의 3차원 배양은 체내 환경에서처럼 세포와 주변 환경의 상호작용이 가능하기 때문에, 특정 장기의 기능이나 질병 상황을 모사할 수 있다. 즉, 암의 전이, 상처회복, 혈관신생, 면역반응 등 다양한 현상에서 관찰되는 세포 및 세포외기질의 상호작용을 실제에 가깝게 관찰하고 분석할 수 있어 약물 평가나 독성평가 분야에서 실제 동물실험과 유사한 결과를 도출할 수 있는 것으로 알려져 있다.Recently, as the results of researches that genes and protein expression are different in two-dimensional cultured cells and three-dimensional cultured cells have been announced, interest in three-dimensional cell culture technology is increasing. The three-dimensional culture of cells can simulate the function or disease state of a specific organ, since the interaction of the cell and the surrounding environment is possible, as in the internal environment. In other words, the interaction of cells and extracellular matrix observed in various phenomena such as cancer metastasis, wound healing, angiogenesis, and immune response can be closely observed and analyzed. Thus, similar to actual animal experiments in the field of drug evaluation and toxicity evaluation Can be derived.
종래의 3차원 배양기술에는 세포를 펠릿(pellet)형태로 뭉친 후 부유 배양하는 방법, 매우 낮은 농도의 세포를 포함한 배양액을 물방울 형태(droplet)로 분주한 다음 배양용기를 뒤집어 액적내에 세포를 부유시켜 배양하는 방법, 미세유체소자와 같은 자동화 배양 시스템을 만들어 3차원 배양을 하는 방법 등이 있다. Conventional three-dimensional culture techniques include a method of culturing cells in the form of pellets followed by suspension culture, a method in which a culture solution containing cells at a very low concentration is dispensed in droplets, the culture container is turned upside down, A method of culturing, and an automated culture system such as a microfluidic device to prepare a three-dimensional culture.
3차원 배양은 2차원 배양에서처럼 세포에 영양을 공급하기 위한 배양액 교환 과정이 필수이다. 그러나 펠릿 배양과 액적 배양은 배양액 교환 과정 중 물리적인 힘에 의한 세포구조체 붕괴가 초래되기 쉽고 시간적인 소요가 커 세포가 외부환경에 오래 노출된다는 단점이 있다. 또한 자동화 배양 시스템은 배양액 교환을 위한 자동화된 프로그램과 장비를 요구하므로 단가가 높다. 따라서 종래기술은 다양한 조건의 3차원 배양 시험을 동시에 수행하는데 한계가 있어, 특히 고속대량스크리닝(high throughput screening)에의 적용은 거의 불가능한 실정이다.Three - dimensional culture is essential for the nutrient supply to cells as in two - dimensional culture. However, pellet culture and droplet cultivation are disadvantageous in that the cell structure collapse due to physical force during the culture exchange process, and the cells are exposed to the external environment for a long period of time because of high time. Automated culture systems also require automated programs and equipment for culture exchange, which is high cost. Therefore, the prior art has a limitation in carrying out a three-dimensional culture test under various conditions at the same time, and it is practically impossible to apply it particularly to high throughput screening.
한편, 최근 세포 배양 장치에 폴리디메틸실록산(Polydimethylsiloxane,, PDMS)이 많이 사용되고 있다. 3차원 PDMS 구조체의 제조는 미소 구조체 제조기술, 즉 미소공정기술에 의해 수행되는데, 그 중 포토리소그래피(Photolithography)가 가장 널리 적용되고 있다. 그러나, 포토리소그래피는 매우 고가의 장비를 사용하기 때문에 제작 단가가 매우 비싸고, 평평한 면에 빛을 쬐어 평면에 패턴을 형성하는 방법을 이용하기 때문에 평평하지 않는 구조체, 예를 들면 곡면을 가지는 구조체를 제작하기에는 어려움이 있다. 또한 제조공정에서 유독한 화학물질을 사용하므로 그 결과물을 생물학분야의 실험에는 사용하기 부적합하며, 환경오염을 유발하고 작업자가 유해 환경에 노출되는 단점이 있다.Recently, polydimethylsiloxane (PDMS) has been widely used in cell culture apparatuses. The fabrication of the three-dimensional PDMS structure is performed by microstructure manufacturing technology, that is, micro-processing technology, among which photolithography is most widely applied. However, since photolithography uses very expensive equipment, the cost of fabrication is very high, and since a method of forming a pattern on a flat surface by applying light to a flat surface is used, a structure having no flat surface, for example, a structure having a curved surface There are difficulties in doing so. In addition, since toxic chemical substances are used in the manufacturing process, the resultant is inadequate to be used for experiments in biological field, and causes environmental pollution and exposes workers to harmful environment.
또 다른 3차원 PDMS 구조체 제조방법에는 소프트리소그래피(Soft lithography)가 있다. 소프트리소그래피는 포토리소그래피와 다르게 폴리디메틸실록산을 사용해서 틀을 만들어 계속 찍어내는 방법이지만, 포토리소그래피의 방식을 통해 틀을 제작하는 방식으로 발전하였기 때문에 상기 포토리소그래피와 동일한 문제가 존재한다. 이러한 단점을 개선하기 위해 다른 방법으로 틀을 만드는 방법들이 제안되고 있지만 제조방법이 까다롭고 여러 종류의 재료를 요구하게 되므로 고가의 비용이 소요되고 있는 실정이다. Another method for manufacturing a three-dimensional PDMS structure is soft lithography. Soft lithography is a method of continuously forming a frame by using polydimethylsiloxane differently from photolithography. However, the same problem as the above photolithography exists because the frame is developed by a method of photolithography. In order to overcome these disadvantages, methods of forming molds by other methods have been suggested, but the manufacturing method is difficult and requires various kinds of materials, so that a high cost is required.
이에, 본 발명자들은 세포의 3차원 배양 시 배양배지 및 시험물질을 용이하게 전달할 수 있고 제작이 용이한 3차원 세포 배양 장치를 개발하고자 예의 노력한 결과, 폴리디메틸실록산 기반 3차원 세포 배양 장치에 상부 표면에 존재하는 배양부 및 배양부와 하부 표면을 연결하는 모세관을 만들고 모세관 내에 젤을 충진하는 경우, 하층의 배양배지 및 시험물질이 모세관 현상에 의하여 배양부에 도달하여 지속적인 물질교환이 이루어지는 것을 확인하고, 본 발명을 완성하게 되었다.
Accordingly, the present inventors have made intensive efforts to develop a three-dimensional cell culture apparatus capable of easily transporting a culture medium and a test substance in a three-dimensional culture of cells and making it easy to produce. As a result, And the capillary tube connecting the culture surface and the lower surface and filling the capillary with the gel, the culture medium and the test substance in the lower layer reach the culture part by the capillary phenomenon, , Thereby completing the present invention.
본 발명의 목적은 폴리디메틸실록산 기반 3차원 세포배양 장치를 제공하는데 있다.
It is an object of the present invention to provide a polydimethylsiloxane-based three-dimensional cell culture apparatus.
상기 목적을 달성하기 위하여, 본 발명은 (a) 상부 표면에 존재하는 배양부; (b) 상기 배양부와 하부 표면을 연결하는 모세관; 및 (c) 모세관을 채우는 충진용 젤을 포함하는 폴리디메틸실록산 기반 3차원 세포 배양 장치를 제공한다.
In order to achieve the above-mentioned object, the present invention provides a method for producing a microorganism, comprising (a) a culture unit present on an upper surface; (b) a capillary connecting the culture section and the lower surface; And (c) a filling gel filling the capillary. The present invention provides a polydimethylsiloxane-based three-dimensional cell culture device comprising:
본 발명에 따르면, 기존의 배양 시스템과의 융합 및 제작이 용이하고, 세포 배양체에 물리적인 자극이 거의 없으며, 고가의 장비 없이 지속적인 물질교환이 가능한 폴리디메틸실록산 기반 3차원 세포 배양 장치를 제공할 수 있으므로, 세포치료제 개발 및 약물 스크리닝 분야에 유용하다.
According to the present invention, it is possible to provide a polydimethylsiloxane-based three-dimensional cell culture device which is easy to be fused and manufactured with an existing culture system, has little physical irritation to the cell culture, and can continuously exchange materials without expensive equipment Therefore, it is useful in the field of cell therapy development and drug screening.
도1은 폴리디메틸실록산 기반 3차원 세포 배양 장치의 모식도이다(A:위에서 내려다 본 구조, B: 단면 구조).
도2는 폴리디메틸실록산 기반 3차원 세포 배양 장치의 제작과정의 예를 나타낸 모식도이다(A: 미세핀 칩을 반응조 내에 놓고 적당량의 PDMS를 붓는 단계; B: PDMS를 굳히는 단계; C: 무수한 구멍이 형서된 PDMS를 수득하고 구멍에 비드(Bead)를 배치하는 단계; D: 반응조로 옮긴 후 적당량의 PDMS를 채우는 단계; E: PDMS를 굳히는 단계; F: 반응조를 뒤집은 다음 하이드로젤을 채워 모세관을 충진하는 단계; G: 하이드로젤을 굳히는 단계; H: 비드를 제거는 단계).
도3은 폴리디메틸실록산 기반 3차원 세포 배양 장치를 종래의 공배양 시스템에 적용하는 방법에 관한 모식도 이다(A: 공배양용 인서트(insert) 적용예시; B: 다층 공배양 적용예시).1 is a schematic view of a polydimethylsiloxane-based three-dimensional cell culture apparatus (A: top view, B: sectional view).
2 is a schematic view showing an example of a process for producing a polydimethylsiloxane-based three-dimensional cell culture apparatus (A: pouring an appropriate amount of PDMS into a reaction vessel, B: hardening PDMS, C: D: Transferring the PDMS to the reaction tank, filling the PDMS with the proper amount, E: Strengthening the PDMS, F: Reversing the reactor, filling the capillary with the hydrogel, G: step of hardening the hydrogel; H: step of removing the beads).
3 is a schematic diagram of a method of applying a polydimethylsiloxane-based three-dimensional cell culture apparatus to a conventional co-cultivation system (A: Example of co-culture insert; B: Multi-layer co-culture application example).
본 발명은 일 관점에서, (a) 상부 표면에 존재하는 배양부; (b) 상기 배양부와 하부 표면을 연결하는 모세관; 및 (c) 모세관을 채우는 충진용 젤을 포함하는 폴리디메틸실록산 기반 3차원 세포 배양 장치에 관한 것이다.According to one aspect of the present invention, there is provided a culture medium comprising (a) a culture medium present on an upper surface; (b) a capillary connecting the culture section and the lower surface; And (c) a filling gel that fills the capillary. The present invention also relates to a polydimethylsiloxane-based three-dimensional cell culture device.
본 발명의 배양부는 3차원 구조를 가지는 것을 특징으로 할 수 있다. The culture unit of the present invention may be characterized by having a three-dimensional structure.
상기 3차원 구조는 단면적이 80 mm2 이하인 것을 특징으로 할 수 있다.The three-dimensional structure may have a cross-sectional area of 80 mm 2 or less.
상기 3차원 구조는 반구형 또는 공동형인 것을 특징으로 할 수 있으나, 이에 한정되는 것은 아니며, 세포가 중력에 의하여 하단으로 모일 수 있도록 하부에 가까울수록 단면적이 줄어드는 것이 바람직하다. The three-dimensional structure may have a hemispherical shape or a hollow shape. However, the present invention is not limited thereto. It is preferable that the cross-sectional area of the three-dimensional structure decreases as the cells are brought closer to the lower part so that the cells can be gathered by gravity.
본 발명의 일 실시예에서는, 정방형의 3차원 구조체로서 구형 비드(Bead)를 몰드(mold)로 사용하여 3차원 세포 배양 장치의 표면에 반구형 배양부를 제조하였다. 본 발명에서는 모세관 구조를 가지는 3차원 세포 배양 장치를 제조하기 위하여 몰드(mold)를 기초로 하는 방법을 사용하였으나 제조방법이 이에 한정되는 것은 아니다.In one embodiment of the present invention, a hemispherical culture unit was prepared on the surface of a three-dimensional cell culture apparatus using a spherical bead as a mold as a square three-dimensional structure. In the present invention, a method based on a mold is used to manufacture a three-dimensional cell culture apparatus having a capillary structure, but the manufacturing method is not limited thereto.
폴리디메틸실록산(Polydimethylsiloxane,, PDMS)은 탄소-산소-수소-규소 화합물로서, 고분자 유기 규소 화합물의 그룹에 속하며 실리콘으로 지칭되는 물질이다. PDMS는 일반적으로 광학적으로 투명하고, 비독성 및 비가연성의 특성을 가져 생물학적, 화학적 시험을 위한 재료로서 널리 이용되고 있는 재료 중 하나이다. 또한 세포독성 실험결과 생체적합성을 지닌 안전한 물질로 밝혀짐에 따라 생체 주입을 위한 소재로도 사용될 수 있다. 특히, PDMS 소자는 산소 투과성이 높기 때문에 세포배양이 안정적으로 가능한 장점이 있으며, 투명하여 고해상도 관찰이 가능하다. Polydimethylsiloxane (PDMS) is a carbon-oxygen-hydrogen-silicon compound that belongs to the group of high molecular organosilicon compounds and is referred to as silicon. PDMS is generally optically transparent, non-toxic and non-flammable, and is one of the widely used materials for biological and chemical tests. As a result of the cytotoxicity test, it has been revealed that it is a safe material having biocompatibility, so that it can be used as a material for bioinjection. Particularly, since the PDMS device has high oxygen permeability, the cell culture can be stably performed, and high-resolution observation is possible with transparency.
본 발명의 다른 실시예에서는 CPU 핀에 폴리디메틸실록산을 부어 다수의 구멍이 존재하는 3차원 세포 배양 장치의 기본 구조를 제조한 다음, 비드(bead)를 얹은 후 다시 폴리디메틸실록산을 부어 반구형 배양부 구조를 추가 하였다.In another embodiment of the present invention, polydimethylsiloxane is poured onto a CPU pin to form a three-dimensional cell culture apparatus having a plurality of pores. Then, a bead is placed on the three-dimensional cell culture apparatus, polydimethylsiloxane is again poured, Structure was added.
본 발명의 모세관은 배양부의 중심에 위치하는 것을 특징으로 할 수 있다.The capillary of the present invention may be characterized by being located at the center of the culture section.
본 발명의 모세관은 단면적이 1 mm2 이하인 것을 특징으로 할 수 있다.The capillary of the present invention can be characterized by a cross-sectional area of 1 mm 2 or less.
본 발명의 또 다른 실시예에서는 지름 0.2mm를 가지는 컴퓨터의 CPU pin을 몰드(mold)로 사용하여, 약 0.03mm2 단면적을 가지는 모세관을 형성하였다. In another embodiment of the present invention, a CPU pin of a computer having a diameter of 0.2 mm is used as a mold to form a capillary having a cross-sectional area of about 0.03 mm 2 .
본 발명의 충진용 젤은 하이드로젤인 것을 특징으로 할 수 있다.The filling gel of the present invention may be a hydrogel.
상기 하이드로젤은 마트리젤(Matrigel), 푸라매트릭스(Puramatrix), 콜라겐(Collagen), 피브린 겔(Fibrin gel), 폴리에틸렌글리콜 디아크릴레이트(PEG-DA), 폴리에틸렌글리콜 디메사크릴레이트(PEG-DMA), 폴리나이팜 (PNIPAM), 폴록세이머 (Poloxamer), 키토산(Chitosan), 아가로스(Agarose), 젤라틴(Gelatin), 히알루론산 (Hyaluronic acid) 또는 알지네이트(Alginate)를 포함하는 것을 특징으로 할 수 있고 폴리에틸렌그리콜 디아크릴레이트인 것이 바람직하다.
The hydrogel may be selected from the group consisting of Matrigel, Puramatrix, Collagen, Fibrin gel, Polyethylene glycol diacrylate (PEG-DA), Polyethylene glycol dimethacrylate (PEG-DMA) (PNIPAM), Poloxamer, Chitosan, Agarose, Gelatin, Hyaluronic acid or Alginate, which can be used in the present invention. Polyethylene glycol diacrylate.
이하, 본 발명을 하기 실시예에에 의해 더욱 상세하게 한다. 이들 실시예는 단지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 범위가 이들 실시예에 국한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.
Hereinafter, the present invention will be described in more detail by way of the following examples. It will be apparent to those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited to these embodiments.
실시예1Example 1 : 모세관 제조: Capillary manufacturing
다수의 모세관을 가지는 배양 장치를 제조하기 위하여 폐기 되는 컴퓨터로부터 중앙처리장치(CPU)를 수득하였다. 핀의 높이는 1.6mm 였고 지름은 0.2mm였다. 수득한 CPU를 수회 세척하고 소독 및 멸균한 후 사용하였다. 또한 폴리디메틸실록산(PDMS; Sylgard 184, Dow Corning, USA) prepolymer와 가교제의 비율을 무게비 10 : 2로 하여 준비하였다. A central processing unit (CPU) was obtained from the computer to be discarded in order to manufacture a culture apparatus having a plurality of capillaries. The height of the pin was 1.6 mm and the diameter was 0.2 mm. The obtained CPU was washed, sterilized and sterilized several times and then used. Further, the ratio of the prepolymer of polydimethylsiloxane (PDMS; Sylgard 184, Dow Corning, USA) to the crosslinking agent was adjusted to a weight ratio of 10: 2.
형틀을 만들기 위한 반응조에 CPU의 핀이 하늘을 향하도록 ?힌 다음, 폴리디메틸실록산(Sylgard 184, Dow Corning, USA)을 부어주었다(도 2A). 구멍이 막히지 않도록 1.5mm 이하로 부어 핀의 끝이 PDMS 용액 표면위로 노출되도록(도 2B) 하였다. PDMS를 굳힌 다음, CPU 핀을 조심스럽게 제거하여 구멍이 형성된 PDMS 젤을 수득하였다.
Polymerization (Sylgard 184, Dow Corning, USA) was poured into the reaction vessel to form the mold so that the pins of the CPU were directed toward the sky (FIG. 2A). The end of the pin was exposed to the surface of the PDMS solution (Fig. 2B) so as not to block the hole. After hardening the PDMS, the CPU pins were carefully removed to obtain a perforated PDMS gel.
실시예2Example 2 : : 배양부Culture unit 제조 Produce
배양부를 제조하기 위하여 상기 실시예1에서 수득한 PDMS 젤에 지름 0.8mm의 금속 비드(bead; 아가미모델링, KR)를 올린 후 가볍게 쳐 각 구멍을 비드가 모두 막도록 하였다. 비드와 구멍을 밀착시키기 위하여 80~100℃에서 반응 시켰으며 3분 후 비드의 반대쪽 구멍에서 음압을 걸어주면서 실온으로 냉각을 시작하였다(도 2C). 비드가 고정된 젤을 새 반응조에 넣고 PDMS 용액을 부어주었다(도 2D). 반구형의 배양부를 만들기 위하여 PDMS를 구의 절반의 위치까지만 채우고(도 2E) 완전히 굳도록 두었다.
In order to manufacture the culture, a bead (gill modeling, KR) having a diameter of 0.8 mm was placed on the PDMS gel obtained in Example 1, and then patched so that the beads were closed. The beads were allowed to react at 80 to 100 ° C to closely contact the beads. After 3 minutes, cooling was started at room temperature while applying a negative pressure to the opposite side of the beads (FIG. 2C). The bead-immobilized gel was placed in a new reaction tank and the PDMS solution was poured (Fig. 2D). To make a hemispherical culture, PDMS was filled to only half of the sphere (Fig. 2E) and allowed to settle completely.
실시예3Example 3 : 모세관 내 젤 : Capillary gel 충진Filling
PDMS 젤 내에 존재하는 비어있는 모세관으로 세포가 유실되지 않는 동시에 영양물질 또는 시험물질이 이동할 수 있도록 모세관 내에 하이드로젤을 충진 하였다. An empty capillary present in the PDMS gel filled the hydrogel in the capillary so that the cells could not be lost and the nutrient or test material could migrate.
상기 실시예2에서 수득한 젤을 뒤집어 비어있는 모세관이 노출시켰다(도 2F). 노출된 구멍으로 10% poly(ethylene glycol) diacrylate(PEGDA; Mw=575; Sigma-Aldrich, USA)에 0.3M Tetramethylethylenediamine(TEMED) 및 0.3M Ammonium persulfate(APS) (sigma-aldrich) 를 첨가한 수용액을 붇고 37℃에서 8분간 굳혀주었다(도 2G).The gel obtained in Example 2 above was turned over to expose an empty capillary (Fig. 2F). An aqueous solution containing 0.3 M Tetramethylethylenediamine (TEMED) and 0.3 M Ammonium persulfate (APS) (Sigma-aldrich) in 10% poly (ethylene glycol) diacrylate (PEGDA; Mw = 575; Sigma-Aldrich, USA) And stiffened at 37 DEG C for 8 minutes (Fig. 2G).
에틸렌글리콜 디아크릴레이트(PEG-DA)이 굳은 후, 젤을 다시 뒤집고 금속 비드를 제거하여 상단에 반구형의 배양부를 가지며, 배양부 중앙에 하이드로젤로 충진된 모세관을 가지는 3차원 세포 배양 장치를 제조하였다(도 2H).After the ethylene glycol diacrylate (PEG-DA) was solidified, the gel was again turned over and the metal beads were removed to prepare a three-dimensional cell culture apparatus having a hemispherical culture unit at the upper part and a capillary filled with hydrogel at the center of the culture unit. (Fig. 2H).
본 발명의 3차원 세포 배양 장치는 다양한 형태로 제조 가능하다. 예를들면 3차원 배양이 불가능한 종래의 공배양 시스템의 인서트(insert)내에 구현함으로써 세포간의 상호작용을 연구할 수 있고(도 3A), 다층의 인서트를 적용한 다중 공배양 또한 가능하다(도 3B).
The three-dimensional cell culture apparatus of the present invention can be manufactured in various forms. For example, it is possible to study intercellular interactions by embodying in an insert of a conventional co-cultivation system which is not capable of three-dimensional culture (Fig. 3A), and multi-co culture with multi-layer insert is also possible (Fig. 3B) .
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의한 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의해 정의된다고 할 것이다.
While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.
Claims (8)
(a) 상부 표면에 존재하는 배양부;
(b) 상기 배양부와 하부 표면을 연결하는 모세관; 및
(c) 모세관을 채우는 충진용 젤.
Polydimethylsiloxane-based three-dimensional cell culture apparatus comprising:
(a) a culture section present on the upper surface;
(b) a capillary connecting the culture section and the lower surface; And
(c) Filling gel filling capillary.
[2] The polydimethylsiloxane-based three-dimensional cell culture apparatus according to claim 1, wherein the culture unit has a three-dimensional structure.
3. The apparatus according to claim 2, wherein the three-dimensional structure has a cross-sectional area of 80 mm 2 or less.
3. The apparatus according to claim 2, wherein the three-dimensional structure is hemispherical or concave.
2. The apparatus according to claim 1, wherein the capillary tube is located at the center of the culture section.
The apparatus of claim 1, wherein the capillary tube has a cross-sectional area of 1 mm 2 or less.
The polydimethylsiloxane-based three-dimensional cell culture apparatus according to claim 1, wherein the filling gel is a hydrogel.
8. The composition of claim 7, wherein the hydrogel is selected from the group consisting of Matrigel, Puramatrix, Collagen, Fibrin gel, Polyethylene glycol diacrylate (PEG-DA), Polyethylene glycol dimethacrylate (PEG-DMA), polynapharma (PNIPAM), poloxamer, chitosan, agarose, gelatin, hyaluronic acid or alginate. Wherein the polydimethylsiloxane-based three-dimensional cell culture apparatus comprises:
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| US11802263B2 (en) | 2017-08-22 | 2023-10-31 | Korea Institute Of Industrial Technology | Culture vessel for three-dimensional cell cultivation and three-dimensional cell co-cultivation method using same |
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| KR20130086016A (en) * | 2013-06-10 | 2013-07-30 | 국립대학법인 울산과학기술대학교 산학협력단 | Cell culture device and method for manufacturing the same |
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