• Title/Summary/Keyword: 3D culture

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Increased SOX2 expression in three-dimensional sphere culture of dental pulp stem cells

  • Seo, Eun Jin;Jang, Il Ho
    • International Journal of Oral Biology
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    • v.45 no.4
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    • pp.197-203
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    • 2020
  • Mesenchymal stem cells in the dental pulp exhibit a tendency for differentiation into various dental lineages and hold great potential as a major conduit for regenerative treatment in dentistry. Although they can be readily isolated from teeth, the exact characteristics of these stem cells have not been fully understood so far. When compared to two-dimensional (2D) cultures, three-dimensional (3D) cultures have the advantage of enriching the stem cell population. Hence, 3D-organoid culture and 3D-sphere culture were applied to dental pulp cells in the current study. Although the establishment of the organoid culture proved unsuccessful, the 3D-sphere culture readily initiated the stable generation of cell aggregates, which continued to grow and could be passaged to the second round. Interestingly, a significant increase in SOX2 expression was detected in the 3D-spheroid culture compared to the 2D culture. These results indicate the enrichment of the stemness-high population in the 3D-sphere culture. Thus, 3D-sphere culture may act as a link between the conventional and 3D-organoid cultures and aid in understanding the characteristics of dental pulp stem cells.

Fabrication and validation study of a 3D tumor cell culture system equipped with bloodvessle-mimik micro-channel (혈관모사 마이크로채널이 장착된 3D 종양 세포 배양 시스템의 제작 및 검증 연구)

  • Park, Jeong-Yeon;Koh, Byum-seok;Kim, Ki-Young;Lee, Dong-Mok;Yoon, Gil-Sang
    • Design & Manufacturing
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    • v.15 no.2
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    • pp.11-16
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    • 2021
  • Recently, three-dimensional (3D) cell culture systems, which are superior to conventional two-dimensional (2D) vascular systems that mimic the in vivo environment, are being actively studied to reproduce drug responses and cell differentiation in organisms. Conventional two-dimensional cell culture methods (scaffold-based and non-scaffold-based) have a limited cell growth rate because the culture cannot supply the culture medium as consistently as microvessels. To solve this problem, we would like to propose a 3D culture system with an environment similar to living cells by continuously supplying the culture medium to the bottom of the 3D cell support. The 3D culture system is a structure in which microvascular structures are combined under a scaffold (agar, collagen, etc.) where cells can settle and grow. First, we have manufactured molds for the formation of four types of microvessel-mimicking chips: width / height ①100 ㎛ / 100 ㎛, ②100 ㎛ / 50 ㎛, ③ 150 ㎛ / 100 ㎛, and ④ 200 ㎛ / 100 ㎛. By injection molding, four types of microfluidic chips were made with GPPS (general purpose polystyrene), and a 100㎛-thick PDMS (polydimethylsiloxane) film was attached to the top of each microfluidic chip. As a result of observing the flow of the culture medium in the microchannel, it was confirmed that when the aspect ratio (height/width) of the microchannel is 1.5 or more, the fluid flows from the inlet to the outlet without a backflow phenomenon. In addition, the culture efficiency experiments of colorectal cancer cells (SW490) were performed in a 3D culture system in which PDMS films with different pore diameters (1/25/45 ㎛) were combined on a microfluidic chip. As a result, it was found that the cell growth rate increased up to 1.3 times and the cell death rate decreased by 71% as a result of the 3D culture system having a hole membrane with a diameter of 10 ㎛ or more compared to the conventional commercial. Based on the results of this study, it is possible to expand and build various 3D cell culture systems that can maximize cell culture efficiency by cell type by adjusting the shape of the microchannel, the size of the film hole, and the flow rate of the inlet.

Analysis of Plasminogen Activators Activity and Three Dimensional (3D) Culture of Endometrial Cells in Pigs (돼지 자궁내막 세포의 3차원 배양과 Plasminogen Activator 활성화 분석)

  • Cha, Hye-Jin;Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.273-280
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    • 2013
  • The aim of this study was to establish a three dimensional (3D) culture system of endometrial cells and to examine the plasminogen activators (PAs) activity in porcine uterine. The 3D culture system in porcine endometrial cells was composed to mixture 3D gel, stromal cells and epithelial cells. The 3D culture system was used to identify normal structure search as uterine tissue and PAs expression in this study. In results, porcine endometrium epithelial cells forming a top monolayer and endometrium stromal cells developed as fibroblast-like within 3D matrix scaffold. Expression of urokinase-type PA (uPA) and tissue-type PA (tPA) were observed during the 3D culture using immunofluorescence. PA activity in 3D-cultured endometrial cells was no significant difference between the tissue type, but 2D culture system were significantly lower than in 3D-cultured endometrial cells (P<0.05). Therefore, basic system and functional aspect of 3D culture could be established with similar system of endometrium tissue. We suggest that this study was assumed applicable as baseline data to investigate mechanism between porcine uterus cells in vitro.

Evaluation of primary hepatocyte function using 2D or 3D culture method for primary rat hepatocytes (Rat Primary Hepatocyte의 2차원 배양과 3차원 배양에 따른 생리 활성능과 대사능에 관한 연구)

  • Lim, Malgum;Kim, Yeongji;Shin, Yurianna;Oh, Keon Bong;Hwang, Seongsoo;Kim, Youngim;Hur, Tai-Young;Ock, Sun A
    • Journal of Embryo Transfer
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    • v.31 no.3
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    • pp.169-177
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    • 2016
  • There is a growing interest in the application of primary hepatocytes for treatment of liver diseases in humans and for drug development. Several studies have focused on long-term survival and di-differentiation blocking of primary hepatocytes in an in vitro culture system. Therefore, the present study also aimed to optimize an in vitro culture system using primary rat hepatocytes. Primary rat hepatocytes from 6-week-old male Crl:CD rats were isolated using a modified two-step collagenase perfusion. Healthy $3.5{\times}10^6$ primary rat hepatocytes were seeded into a 2 dimensional (2D) culture in a 25T culture flask coated with collagen type I or into a 3D culture in a 125-ml spinner flask for 7 days. Production of plasma protein (ALB and TF), apoptosis (BAX and BCL2), and CYP (CYP3A1) related genes were compared between the 2D and 3D culture systems. The 3D culture system had an advantage over the 2D system because of the relatively high expression of ALB and low expression of BAX in the 3D system. However, the level of CYP3A1 did not improve in the 3D culture with and without the presence of a dexamethasone inducer. Therefore, 3D culture has an advantage for albumin production and primary rat hepatocyte survivability, but a low expression of CYP3A1 indicated that primary rat hepatocytes require a high-density culture for stress reduction by continuous flow.

Microbial Basis for Enhanced Degradation of the Fumigant 1,3-Dichloropropene (1,3-D) in Soil

  • Chung, Keun-Yook
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.10a
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    • pp.125-139
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    • 2000
  • The differential enhanced degradation of cis- and trans-1,3-D was observed in the previous two studies performed by Ou et al. (1995) and especially Chung et al. (1999). This study was initiated to investigate the involvement of microorganisms in the differential enhanced degradation of the chemicals. As expected, microorganisms were responsible for the enhanced degradation of the chemicals. A mixed bacterial culture capable of degrading 1,3-D was isolated from an enhanced soil sample collected from a site treated with 1,3-D. Similar to the enhanced soil, the mixed culture degraded trans-1,3-D faster than cis-1,3-D. This mixed culture could not utilize cis- and trans-1,3-D as a sole source of carbon for growth. Rather, a variety of second substrates were evaluated to stimulate the differential enhanced degradation of the two isomers. As a result, the mixed culture degraded cis- and trans-1,3-D only in the presence of a suitable second substrate. Second substrates that had the capacity to stimulate the degradation included soil leachate, tryptone, tryptophan, and alanine. Other substrates tested, including soil extract, glucose, yeast extract, and indole (ailed to stimulate the degradation of the two isomers. Therefore, it appeared that the degradation of cis- and trans-1,3-D was a cometabolic process. The mixed culture was composed of four morphologically distinctive bacterial colonies.

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A Study on 3D Culture Character Modeling for Curriculum (창업을 위한 3D문화캐릭터 모델링 교육과정에 관한 연구)

  • Park, Hea-sook
    • Proceedings of the Korean Society of Computer Information Conference
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    • 2018.01a
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    • pp.211-212
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    • 2018
  • 본 논문에서는 문화콘텐츠 캐릭터를 3D모델링 도구를 이용하여 디자인하고 3D 프린팅 기술을 접목하여 출력하여 판매하고자 하는 창업에 대한 교육과정을 제시하고자 한다. 이를 위해 첫 단계로서 3D 모델링 도구의 전반적인 특성을 살펴보고 3D 캐릭터 모델링 교육을 위한 교육과정으로 어떠한 교과목들이 제시되어야 하는지 제시해보고자 한다.

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A Study of the Visual Expressions of Traditional Culture in the 3D Animation Chang'an

  • Lei Xu;Jeanhun Chung
    • International journal of advanced smart convergence
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    • v.13 no.2
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    • pp.136-141
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    • 2024
  • Chang'an (Chinese: 长安三万里), also known as 30,000 Miles from Chang'an, is a 2023 Chinese 3D animated historical drama film directed by Xie Junwei and Zou Jing.This thesis aims to explore the visual expression of traditional culture in the 3D animated film Chang'an as an example to reveal the reasons for the success of this type of film. The study analyses in detail the design of the character models and costumes, as well as the use of the traditional landscape painting technique of 'white space' in the composition of the screen from the visual aspect. Through the analysis of character design and screen composition, the thesis concludes that the success of Chang'an lies in its elaborate visual design and clever use of traditional culture, which makes it a 3D animation film with both artistic and commercial values. Finally, the thesis concludes that the production of a successful 3D animation film needs to combine the visual elements of 3D animation with traditional culture in order to win audience recognition and achieve commercial success.

Phytochemical Constituents of Acanthopanax senticosus (Rupr. & Maxim.) Harms Stem

  • Ryu, Ji-Young;Son, Dong-Wook;Kang, Jung-Il;Lee, Sang-Yun;Kim, Hyun-Su;Shin, Kuk-Hyun;Lee, Sang-Hyun
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.4
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    • pp.306-310
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    • 2003
  • Five constituents were isolated from the stem of Acanthopanax senticosus. Their structures were elucidated as (-)-sesamin (1), iso-fraxidin (2), 5-hydroxymethylfurfural (3), syringin (4) and acanthoside D (5) by spectral analysis. Among these compounds, 5-hydroxymethylfurfural (3) was isolated for the first time from this plant.

Actinomycins에 의한 Adenosine Deaminase의 억제

  • 김경자;조성진
    • Microbiology and Biotechnology Letters
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    • v.24 no.3
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    • pp.380-383
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    • 1996
  • Adenosine deaminase inhibitor was extracted from culture broth of Streptomyces sp. strain V-8 with ethylacetate. The ethylacetate extract showed the characteristic UV absorption spectrum of actinomycins at 440-450 nm. The ethylacetate extract was compared with respect to inhibitory behavior against adenosine deaminase from calf intestinal mucosa with actinomycin D, -C complex and actinomycin V. The Ki values for actnomycin D, -C complex, and actinomycin V against adenosine deaminase were determined to be 9.9 $\times$ 10$^{-6}$ M, 9.6 $\times$ 10$^{-6}$ M and 9.3 $\times$ 10$^{-6}$ M, respectively. The Ki value for the ethylacetate extract of culture broth against adenosine deaminase was determined to be 5.7 $\times$ 10$^{-6}$ M. The kinetic parameters of actinomycin D, -C complex, -V and ethylacetate extract of culture broth for adenosine deaminase were as follows:I$_{50}$ = 1.5 $\times$ 10$^{-5}$ M (actinomycin D), 2.7 $\times$ 10$^{-5}$ M (actinomycin C complex), 3.5 $\times$ 10$^{-5}$ M (actinomycin V), 8.9 $\times$ 10$^{-6}$ M (ethylacetate extract of culture broth). The adenosine deaminase was inhibited noncompetitively by ethylacetate extract of culture broth as well as by actinomycin D, -C complex and actinomycin V.

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Effect of Different Volume of Microdrop Culture on B6D2F1 Mice Oogenesis (배양액 용량이 B6D2F1 마우스 배아발생능력에 미치는 영향)

  • Yoo, Chang-Seok;Park, Kee Sang;Seo, Byoung Boo
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.27-32
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    • 2016
  • This study was conducted to evaluate the effects of different volume ($100{\mu}l$ vs. 2 ml) of microdrop culture on B6D2F1 mice oogenesis. In the present study, B6D2F1/CrljOri $F_1$ mice were utilized in order to maximize oogenesis. Also we used TCM-199, Dulbecco's medified Eagle's medium (DMEM), embryo culture medium (Fertilization medium, Cleavage medium, Blastocyst medium), G series medium and One step medium. Blastulation rate was not different between groups ($58.4{\pm}2.9%$ vs. $61.2{\pm}4.8%$). Zona hatched rate ($38{\pm}15.4%$ vs. $27{\pm}3.4%$) and attached rate ($55{\pm}13.9%$ vs. $46{\pm}3.9%$) did not differ by the volume of culture media. Total cell numbers ($59.8{\pm}9.7$ vs. $70.3{\pm}8.7$), ICM cell numbers ($15.8{\pm}0.6$ vs. $16.8{\pm}1.5$), TE cell numbers ($44.0{\pm}9.7$ vs. $53.6{\pm}7.3$), % ICM ($26.4{\pm}2.9%$ vs. $23.8{\pm}3.3%$) and ICM:TE ratio ($1:2.8{\pm}0.4$ vs. $1:3.2{\pm}0.6$) were not different between groups (i.e., $100{\mu}l$ vs. 2 ml). These results show that the capacity of the culture medium did not effect the cell numbers of B6D2F1 mice blastocysts. In summary, these results can provide fundamental data to maximize culture condition for in vitro fertilization on B6D2F1 mice.