• Title/Summary/Keyword: microwells

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The effects of ascorbic acid on the morphology and cell proliferation of three-dimensional stem cell spheroids (아스코르브산의 3차원 줄기세포 배양체에 대한 증식 및 분화 효과 분석)

  • Lee, Hyunjin;Yeo, Seong-Il;Park, Jun-Beom
    • The Journal of the Korean dental association
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    • v.55 no.9
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    • pp.592-603
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    • 2017
  • Purpose: The effects of various concentrations of ascorbic acid on stem cell spheroids derived from intraoral areas are not known yet. Thus, the purpose of this study is to evaluate the effects of different concentrations of ascorbic acid on the morphology and cellular viability of stem cell spheroids derived from the gingival tissues. Materials and Methods: Stem cells were plated onto silicon elastomer-based concave microwells and grown in the presence of ascorbic acid at concentrations ranging from 0.003% to 0.3%. The morphology of the cells was viewed under an inverted microscope at day 1, 2, 3 and 5. Qualitative live/dead assay and quantitative cellular viability using Cell Counting Kit-8 were performed on day 2 and day 5. Results: Gingiva-derived stem cells formed spheroids irrespective of ascorbic acid concentration in silicon elastomer-based concave microwells. Increase in the diameter of spheroid were seen with higher concentrations of ascorbic acid. Higher cellular viability was seen in higher concentrations of ascorbic acid. Conclusion: Within the experimental setting, the application of ascorbic acid on stem-cell spheroids produced an increase in the size and higher viability with higher dosage. It can be suggested ascorbic acid be applied with stem cell spheroids for tissue engineering purposes.

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Monocyte Attachment and Migration through Collagen IV in Diabetes Mellitus

  • Kostidou, Elena;Koliakos, George;Paletas, Konstantinos;Kaloyianni, Martha
    • Molecules and Cells
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    • v.25 no.3
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    • pp.452-456
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    • 2008
  • The interactions between monocytes and extracellular matrix proteins have been implicated in atherosclerosis pathophysiology. In the present study we evaluated monocyte attachment and migration through oxidized and non-oxidized collagen IV. Monocyte attachment was tested on microwells coated with either native or oxidized collagen IV. Monocyte migration through collagen IV was examined on transwells. Monocytes derived from patients with diabetes mellitus showed an increased ability to attach and migrate through collagen IV as compared to those derived from healthy volunteers. Moreover, control monocytes attached to oxidized collagen at a higher degree, while they migrated through oxidized collagen at a lower degree, as compared to the native protein. Our results also showed the involvement of the alpha2 integrin subunit in the above phenomena suggesting a modified interaction between monocytes and collagen IV in diabetes mellitus.

Fabrication of 2-D Microwell Arrays for Micro ELISA System (초소형 면역 분석기용 이차원적 배열형 마이크로 우물 제작)

  • Lee, Kook-Nyung;Ahn, Si-Hong;Park, Jae-Hyung;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 1998.07g
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    • pp.2556-2558
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    • 1998
  • 2-D microwell arrays for micro ELISA (Enzyme-Linked Immuno Solvent Assay) system were fabricated using micromachining technology. The materials for the bottom plate, top plate and sidewall of the microwell were used a #7740 glass, gold and silicon respectively considering bio-compatibility and easy fabrication. Cylindrical or groove shape microwells, which have about $100{\mu}m$ depth and $50{\sim}500{\mu}m$ diameter or width, were arrayed. The fabricated microwell array can be applied to the essential part of a biochip when surface modification is made to immobilize cells or biomolecules on the microwell bottom.

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Engineered microdevices for single cell immunological assay

  • Choi, Jong-Hoon
    • Interdisciplinary Bio Central
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    • v.2 no.2
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    • pp.1.1-1.8
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    • 2010
  • Microdevices have been used as effective experimental tools for the rapid and multiplexed analysis of individual cells in single-cell assays. Technological advances for miniaturizing such systems and the optimization of delicate controls in micron-sized space homing cells have motivated many researchers from diverse fields (e.g., cancer research, stem cell research, therapeutic agent development, etc.) to employ microtools in their scientific research. Microtools allow high-throughput, multiplexed analysis of single cells, and they are not limited by the lack of large samples. These characteristics may significantly benefit the study of immune cells, where the number of cells available for testing is usually limited. In this review, I present an overview of several microtools that are currently available for single-cell analyses in two popular formats: microarrays and microfluidic microdevices. Then, I discuss the potential to study human immunology on the single-cell level, and I highlight several recent examples of immunoassays performed with single-cell microdevice assays. Finally, I discuss the outlook for the development of optimized assay platforms to study human immune cells. The development and application of microdevices for studies on single immune cells presents novel opportunities for the qualitative and quantitative characterization of immune cells and may lead to a comprehensive understanding of fundamental aspects of human immunology.

Design and Fabrication of Mold Insert for Injection Molding of Microfluidic tab-on-a-chip for Detection of Agglutination (응집반응 검출을 위한 미세 유체 Lab on a chip의 사출성형 금형 인서트의 디자인 및 제작)

  • Choi, Sung-Hwan;Kim, Dong-Sung;Kwon, Tai-Hun
    • Transactions of Materials Processing
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    • v.15 no.9 s.90
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    • pp.667-672
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    • 2006
  • Agglutination is one of the most commonly employed reactions in clinical diagnosis. In this paper, we have designed and fabricated nickel mold insert for injection molding of a microfluidic lab-on-a-chip for the purpose of the efficient detection of agglutination. In the presented microfluidic lab-on-a-chip, two inlets for sample blood and reagent, flow guiding microchannels, improved serpentine laminating micromixer(ISLM) and reaction microwells are fully integrated. The ISLM, recently developed by our group, can highly improve mixing of the sample blood and reagent in the microchannel, thereby enhancing reaction of agglutinogens and agglutinins. The reaction microwell was designed to contain large volume of about $25{\mu}l$ of the mixture of sample blood and reagent. The result of agglutination in the reaction microwell could be determined by means of the level of the light transmission. To achieve the cost-effectiveness, the microfluidic lab-on-a-chip was realized by the injection molding of COC(cyclic olefin copolymer) and thermal bonding of two injection molded COC substrates. To define microfeatures in the microfluidic lab-on-a-chip precisely, the nickel mold inserts of lab-on-a-chip for the injection molding were fabricated by combining the UV photolithography with a negative photoresist SU-8 and the nickel electroplating process. The microfluidic lab-on-a-chip developed in this study could be applied to various clinical diagnosis based on agglutination.

Simple Fabrication of Adipocyte Cell Chip Using Micropatterning (미세접촉인쇄법을 이용한 지방세포 칩 제작)

  • Kim, Gi Yong;Jeong, Heon-Ho;Lee, Chang-Soo;Roh, Changhyun
    • Korean Chemical Engineering Research
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    • v.54 no.2
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    • pp.223-228
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    • 2016
  • In this study, we described a simple and facile method to generate uniform microwells poly(dimethyl siloxane) (PDMS) microstamps through micro-molding for efficient, rapid and reliable cell patterning of adipocyte differentiation. In contrast to the conventional methods, the microstamp technologies are low expensive, non-toxic, and using a small amount of solution. Recently, Orlistat known as tetrahydrolipstatin is a prescription drug designed to treat obesity which is used to aid in weight loss and help to reduce overweight obesity. Here, 3T3-L1 cells were treated under various concentration manners of Orlistat $0.2{\mu}M{\sim}5.0{\mu}M$. and it was confirmed maximum 26.5% inhibition activity compared to control. Thus, we elucidated this platform can be used for the real-time analyzing of cell proliferation, adipocyte differentiation for evaluation of anti-obesity agents on cell chip. Furthermore, we except that this platform technology designed here might be readily be expanded to discover a wider variety of anti-obesity agents.

Enhancement of preimplantation mouse embryo development with optimized in vitro culture dish via stabilization of medium osmolarity

  • Hyejin Yoon;Jongwoo Lee;Inyoung Kang;Kyoo Wan Choi;Jaewang Lee;Jin Hyun Jun
    • Clinical and Experimental Reproductive Medicine
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    • v.50 no.4
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    • pp.244-252
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    • 2023
  • Objective: We evaluated the efficacy of the newly developed optimized in vitro culture (OIVC) dish for cultivating preimplantation mouse embryos. This dish minimizes the need for mineral oil and incorporates microwells, providing a stable culture environment and enabling independent monitoring of individual embryos. Methods: Mouse pronuclear (PN) zygotes and two-cell-stage embryos were collected at 18 and 46 hours after human chorionic gonadotropin injection, respectively. These were cultured for 120 hours using potassium simplex optimized medium (KSOM) to reach the blastocyst stage. The embryos were randomly allocated into three groups, each cultured in one of three dishes: a 60-mm culture dish, a microdrop dish, and an OIVC dish that we developed. Results: The OIVC dish effectively maintained the osmolarity of the KSOM culture medium over a 5-day period using only 2 mL of mineral oil. This contrasts with the significant osmolarity increase observed in the 60-mm culture dish. Additionally, the OIVC dish exhibited higher blastulation rates from two-cell embryos (100%) relative to the other dish types. Moreover, blastocysts derived from both PN zygotes and two-cell embryos in the OIVC dish group demonstrated significantly elevated mean cell numbers. Conclusion: Use of the OIVC dish markedly increased the number of cells in blastocysts derived from the in vitro culture of preimplantation mouse embryos. The capacity of this dish to maintain medium osmolarity with minimal mineral oil usage represents a breakthrough that may advance embryo culture techniques for various mammals, including human in vitro fertilization and embryo transfer programs.