• Title/Summary/Keyword: Dynamic cell culture

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Design of an Actuator Using Electro-active Polymer (EAP) Actuator with Composite Electrodes (복합재료 전극을 가진 전기활성고분자 구동기의 설계)

  • Kim, Dong-Uk;Chang, Seung-Hwan
    • Composites Research
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    • v.32 no.5
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    • pp.211-215
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    • 2019
  • The cell culture process under in vitro condition is much different from the actual human body environment. Therefore, in order to precisely simulate the human body environment, a dynamic cell culture device capable of delivering mechanical stimulation to cells is essential. However, conventional dynamic cell culture devices require relatively complicated devices such as tubes, pumps, and motors, and the mechanical stimuli delivered is also simple. In this study, an electro-active polymer actuator as a driving component is introduced to design simply driven dynamic cell culture device without complicated components. The device is capable of delivering relatively complex mechanical stimuli to the cells.

Comparative Study of Seeding and Culture Methods to Vascular Smooth Muscle Cells on Biodegradable Scaffold

  • Kim, Dong-Ik;Park, Hee-Jung;Eo, Hyun-Seoun;Suh, Soo-Won;Hong, Ji-Hee;Lee, Min-Jae;Kim, Jong-Sung;Jang, In-Sung;Kim, Byung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.707-714
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    • 2004
  • How to improve the cell culture method on scaffolds is important in the tissue engineering fileld. In this study, we optimized seeding and culture methods to vascular smooth muscle cells (VSMCs) on biodegradable polymer scaffold. The primary culture of VSMCs obtained from canine external jugular vein was accomplished by applying the explant-derived method. The primary cultured VSMCs were seeded into scaffolds and then cultured by using various different methods; static or dynamic seeding, static or dynamic culture. The difference in proliferative response of VSMCs was analyzed with an alamar blue assay. Cell-polymer construct was examined by histochemical method and scanning electron microscopy. Mesh type scaffold ($10 \times 10 \times0.4 mm$) was made of polyglycolic acid (PGA) suture thread. The PGA mesh type scaffold was 45% in porosity, and 0.03 g in weight. The primary cultured VSMCs were confirmed with immunohistochemical staining using monoclonal anti-$\alpha$-smooth muscle actin. The density and distribution of proliferated VSMCs within the scaffold and cellular adherence on the surface of the scaffold showed better results in the static seeding condition than in the dynamic condition. Under the same condition of seeding method as the static condition, the dynamic culture condition showed enhanced proliferation rates of the VSMCs when compared to the static culture condition. In conclusion, to improve the VSMCs proliferation in vitro, static seeding is better than the dynamic condition. In the culture condition, however, culture under the dynamic status is better than the static condition. This was a pilot study to manufacture artificial vascular vessel by tissue engineering.

Effects of Mechanical Stimulation for MC3T3-E1 Cells using Bioreactor (바이오리액터를 이용한 MC3T3-E1 세포의 기계적 자극에 대한 영향)

  • Lee, In-Hwan;Park, Jeong-Hun;Lee, Seung-Jae;Cho, Dong-Woo;Kang, Sang-Sun
    • Proceedings of the KSME Conference
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    • 2008.11a
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    • pp.1411-1414
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    • 2008
  • It is reported that mechanical stimulation takes a role in improving cell growth in skeletal system. And various research groups have showed that developed bioreactor to stimulate cell-seeded and threedimensional scaffold. In this study, we designed a custom-made bioreactor capable of applying controlled compression to cell-seeded agarose gel. This device consisted of a circulation system and compression system. In circular system, culture chamber was sealed for prohibiting contamination and media solution was circulated by pump. In compression system, mechanical stimuli were controlled by LabVIEW software and mechanical transfer system. Cell-encapsulated agarose gels were cultured for up to 7 days. There were significant differences between the number of cells grown in dynamic cell culture and in static cell culture from 3 days to 7 days.

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Viable Alternatives to in vivo Tests for Evaluating the Toxicity of Engineered Carbon Nanotubes

  • Kwon, Soon-Jo;Eo, Soo-Mi
    • Journal of Environmental Health Sciences
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    • v.38 no.1
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    • pp.1-7
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    • 2012
  • Carbon nanotubes (CNTs) stand at the frontier of nanotechnology and are destined to stimulate the next industrial revolution. Rapid increase in their production and use in the technology industry have led to concerns over the effects of CNT on human health and the environment. The prominent use of CNTs in biomedical applications also increases the possibility of human exposure, while properties such as their high aspect ratio (fiber-like shape) and large surface area raise safety concerns for human health if exposure does occur. It is crucial to develop viable alternatives to in vivo tests in order to evaluate the toxicity of engineered CNTs and develop validated experimental models capable of identifying CNTs' toxic effects and predicting their level of toxicity in the human respiratory system. Human lung epithelial cells serve as a barrier at the interface between the surrounding air and lung tissues in response to exogenous particles such as air-pollutants, including CNTs. Monolayer culture of the key individual cell types has provided abundant fundamental information on the response of these cells to external perturbations. However, such systems are limited by the absence of cell-cell interactions and their dynamic nature, which are both present in vivo. In this review, we suggested two viable alternatives to in vivo tests to evaluate the health risk of human exposure to CNTs.

Effects of dynamic oxygen concentrations on the development of mouse pre- and peri-implantation embryos using a double-channel gas supply incubator system

  • Lee, Seung-Chan;Seo, Ho-Chul;Lee, Jaewang;Jun, Jin Hyun;Choi, Kyoo Wan
    • Clinical and Experimental Reproductive Medicine
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    • v.46 no.4
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    • pp.189-196
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    • 2019
  • Objective: We aimed to evaluate the effects of different oxygen conditions (20% [high O2], 5% [low O2] and 5% decreased to 2% [dynamic O2]) on mouse pre- and peri-implantation development using a novel double-channel gas supply (DCGS) incubator (CNC Biotech Inc.) to alter the oxygen concentration during in vitro culture. Methods: The high-O2 and low-O2 groups were cultured from the one-cell to the blastocyst stage under 20% and 5% oxygen concentrations, respectively. In the dynamic-O2 group, mouse embryos were cultured from the one-cell to the morula stage under 5% O2 for 3 days, followed by culture under 2% O2 to the blastocyst stage. To evaluate peri-implantation development, the blastocysts from the three groups were individually transferred to a fibronectin-coated dish and cultured to the outgrowth stage in droplets. Results: The blastocyst formation rate was significantly higher in the low-O2 and dynamic-O2 groups than in the high-O2 group. The total cell number was significantly higher in the dynamic-O2 group than in the low-O2 and high-O2 groups. Additionally, the apoptotic index was significantly lower in the low-O2 and dynamic-O2 groups than in the high-O2 group. The trophoblast outgrowth rate and spread area were significantly higher in the low-O2 and dynamic-O2 groups than in the high-O2 group. Conclusion: Our results showed that a dynamic oxygen concentration (decreasing from 5% to 2%) had beneficial effects on mouse pre- and peri-implantation development. Optimized, dynamic changing of oxygen concentrations using the novel DCGS incubator could improve the developmental competence of in vitro cultured embryos in a human in vitro fertilization and embryo transfer program.

Development of Bioreactor by Rapid Prototyping Technology (쾌속 조형 기술을 이용한 바이오리액티의 개발)

  • Park, Jeong-Hun;Lee, Seung-Jae;Lee, In-Hwan;Cho, Dong-Woo;Rhie, Jong-Won
    • Journal of the Korean Society for Precision Engineering
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    • v.26 no.3
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    • pp.137-143
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    • 2009
  • It has been reported that mechanical stimulation takes a role in improving eel/ growth in skeletal system. Various research groups have been showed their own bioreactors which stimulate cell-seed three-dimensional scaffold. In this study, we hypothesized that the various conditions of mechanical stimulation would affect cell growth and proliferation. To prove our hypothesis, we designed a custom-made bioreactor capable of applying controlled compression to cell-encapsulated scaffolds. This device consisted of a circulation system and a compression system. Each parts of the bioreactor was fabricated using the rapid prototyping technology By using the rapid prototyping technology, we can modify and improve the bioreactor very rapidly For dynamic cell-culture, cell-encapsulated agarose gel was fabricated in 2% concentration. We performed dynamic cell-culture using this agarose gel and developed bioreactor in 3 days.

The Effect of Intermittent Compressive Loading to Growth of Pre-osteoblast Cells (간헐적인 압축하중이 조골세포주 성장에 미치는 영향)

  • Choi, Sung-Kyu;Park, Jeong-Hun;Lee, Seung-Jae;Lee, In-Hwan;Kang, Sang-Sun
    • Journal of the Korean Society for Precision Engineering
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    • v.27 no.2
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    • pp.153-159
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    • 2010
  • Recently, it has been reported that mechanical stimulation takes a role in improving cell growth. Also, became generally known that skeletal system as bone or cartilage tissues take influence of compression loading. In this study, we fabricated a custom-made bioreactor and analyzed that conditions of compressive loading would influence cell growth. To compare the effect of intermittent compressive loading on cell-encapsulated agarose scaffold, we cultured preosteoblast cell (MC3T3-E1 cells) statically and dynamically. And dynamic culture conditions were produced by changing parameters such as the iteration time and interval delay time. Also, cellencapsulated agarose scaffold were subjected to 10 % dynamic compressive strain at 1㎐ frequency for 7 days. After cell culture, cell proliferation was assessed with PI stain assay for fluorescence images and flow cytometry (FACS).

Non-invasive Methods for Determination of Cellular Growth in Podophyllum hexandrum Suspension Cultures

  • Chattopadhyay, Saurabh;Bisaria, V.S.;Scheper, T.;Srivastava, A.K.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.6
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    • pp.331-334
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    • 2002
  • Culture conductivity and on-line NADH fluorescence were used to measure cellular growth in plant cell suspension cultures of Podophyllum hexandrum. An inverse correlation between dry cell weight and medium conductivity was observed during shake flask cultivation. A linear relationship between dry cell weight and culture NADH fluorescence was obtained during the exponential phase of batch cultivation In a bioreactor under the pH stat (pH 6) conditions. It was observed that conductivity measurement were suitable for biomass characterisation under highly dynamic uncontrolled shake flask cultivation conditions. However, if the acid/alkali feeding is done for pH control the conductivity measurement could not be applied. On the other hand the NADH fluorescence measurement allowed online-in situ biomass monitoring of rather heterogenous plant cell suspension cultures in bioreactor even under the most desirable pH stat conditions.

On-Line Estimation of Cell Growth from Agitation Speed in DO-Stat Culture of a Filamentous Microorganism, Agaricus blazei

  • Na, Jeong-Geol;Kim, Hyun-Han;Chang, Yong-Keun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.571-575
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    • 2005
  • A simple, but effective on-line method for estimating the mycelial cell mass concentration from agitation speed data, a most readily-available process variable, has been developed for DO-stat cultures of Agaricus blazei. The dynamic change of dissolved oxygen concentration (DOC) in the initial transient period and the change in yield were considered in the development of the estimation algorithm or estimator. Parameters in the estimation algorithm were calculated from the agitation speed data at 20% of DOC. The proposed estimator could accurately predict the cell mass concentration regardless of DOC levels in the tested range of $10{\sim}40%$, showing a good extrapolation capability.

Reconstruction of Rabbit Corneal Epithelium using Lyophilized Amniotic Membrane and Dynamic Culture Method (동결건조 양막과 동적배양법을 이용한 토끼 각막 상피층의 재구성)

  • Ahn, Jae-Il;Jang, In-Keun;Shin, Youn-Ho;Seo, Young-Kwon;Yoon, Hee-Hun;Yoon, Mun-Young;Kim, Jae-Chang;Song, Kye-Yong;Lee, Hee-Gu;Yang, Eun-Kyung;Kim, Ki-Ho;Park, Jung-Keug
    • KSBB Journal
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    • v.20 no.4
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    • pp.305-310
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    • 2005
  • Reconstruction of rabbit corneal epithelium was performed through dynamic culture method using self-manufactured amniotic membrane supporter and lyophilized amniotic membrane. Rabbit corneal epithelial cells were cultured and cryopreserved after isolation from limbus, and the cells could be proliferated by passage number 10. The basal layer was well formed, and the epithelium layer was constructed tightly by the increase of cell proliferation and differentiation by dynamic culture method than static culture. Thus, the reconstruction of the corneal epithelium using lyophilized amniotic membrane is considered to be a good in vitro model for transplantation of corneal epithelium to patients with a severely damaged cornea.