• Title/Summary/Keyword: 3D cell culture

Search Result 524, Processing Time 0.033 seconds

Electrohydrodynamic Jet Process for Pore-Structure-Controlled 3D Fibrous Architecture As a Tissue Regenerative Material: Fabrication and Cellular Activities

  • Kim, Minseong;Lee, Hyeongjin;Kim, GeunHyung
    • Proceedings of the Korean Institute of Surface Engineering Conference
    • /
    • 2017.05a
    • /
    • pp.134.1-134.1
    • /
    • 2017
  • In this study, we propose a new scaffold fabrication method, "direct electro-hydrodynamic jet process," using the initial jet of an electrospinning process and ethanol media as a target. The fabricated threedimensional (3D) fibrous structure was configured with multilayered microsized struts consisting of randomly entangled micro/nanofibrous architecture, similar to that of native extracellular matrixes. The fabrication of the structure was highly dependent on various processing parameters, such as the surface tension of the target media, and the flow rate and weight fraction of the polymer solution. As a tissue regenerative material, the 3D fibrous scaffold was cultured with preosteoblasts to observe the initial cellular activities in comparison with a solid-freeform fabricated 3D scaffold sharing a similar structural geometry. The cell-culture results showed that the newly developed scaffold provided outstanding microcellular environmental conditions to the seeded cells (about 3.5-fold better initial cell attachment and 2.1-fold better cell proliferation).

  • PDF

Factors influencing efficiency of somatic embryogenesis of Gentiana kurroo (Royle) cell suspension

  • Fiuk, Agnieszka;Rybczynski, Jan J.
    • Plant Biotechnology Reports
    • /
    • v.2 no.1
    • /
    • pp.33-39
    • /
    • 2008
  • In this paper, we would like to show unexpected morphogenic potential of cell suspensions derived from seedling explants of Gentiana kurroo (Royle). Suspension cultures were established with the use of embryogenic callus derived from seedling explants (root, hypocotyl and cotyledons). Proembryogenic mass proliferated in liquid MS medium supplemented with $0.5mg\;l^{-1}$ 2,4-D and $1.0mg\;l^{-1}$ Kin. The highest growth coefficient was achieved for root derived cell suspensions. The microscopic analysis showed differences in aggregate structure depending on their size. To assess the embryogenic capability of the particular culture, 100 mg of cell aggregates was implanted on MS agar medium supplemented with Kin ($0.0-2.0mg\;l^{-1}$), $GA_3$ ($0.0-2.0mg\;l^{-1}$) and AS ($80.0mg\;l^{-1}$). The highest number of somatic embryos was obtained for cotyledon-derived cell suspension on $GA_3$-free medium, but the best morphological quality of embryos was observed in the presence of $0.5-1.0mg\;l^{-1}$ Kin, $0.5mg\;l^{-1}$ $GA_3$ and $80.0mg\;l^{-1}$ AS. The morphogenic competence of cultures also depended on the size of the aggregate fraction and was lower when size of aggregates decreased. Flow cytometry analysis reveled luck of uniformity of regenerants derived from hypocotyl suspension and 100% of uniformity for cotyledon suspension.

Plant Regeneration from Cell Suspension Culture Using Leaf Callus in Actinidia deliciosa X A. arguta Clone 118 (양다래X다래 클론 118의 엽조직 캘러스를 이용한 세포 현탁배양으로부터 식물체 유도)

  • Kim Yong-Wook;Moon Heung-Kyu
    • Journal of Plant Biotechnology
    • /
    • v.32 no.4
    • /
    • pp.287-292
    • /
    • 2005
  • Calli were induced by culturing the leaf segment of Actinidia deliciosa ${\times}$ A. arguta clone 118 on MS medium supplemented with 0.5 mg/L 2,4-D, 0.1 mg/L NAA and 0.05 mg/L BA for 8 weeks in light condition. The induced calli were inoculated in liquid MS medium containing 0.5 mg/L 2,4-D, 0.1 mg/L NAA, 0.05 mg/L BA and 3% sucrose to establish cell suspension culture. The cells at the exponential stage and the stationary stage could be observed between 5-11 days and after that 12 days in culture, respectively. The fresh weight of callus induced from the suspended cells did not vary much among the media containing eight different combinations of plant growth regulators tested. The highest frequency of shoot induction (88.3%) was observed in MS medium containing 2.0 mg/L zeatin. Either BA or zeatin mixed with thidiazuron (TDZ) seemed to be effective in shoot induction. The induced shoots were transferred to MS medium containing 0.2 mg/L zeatin for further shoot growth. And then the shoots were transferred to Standardi (ST) medium containing 1.0 mg/L indolebutyric acid (IBA) for rooting. Plantlets could be obtained through cell suspension culture of Actinidia deliciosa ${\times}$ A. arguta clone 118.

Three-Dimensional Skin Tissue Printing with Human Skin Cell Lines and Mouse Skin-Derived Epidermal and Dermal Cells

  • Jin, Soojung;Oh, You Na;Son, Yu Ri;Kwon, Boguen;Park, Jung-ha;Gang, Min jeong;Kim, Byung Woo;Kwon, Hyun Ju
    • Journal of Microbiology and Biotechnology
    • /
    • v.32 no.2
    • /
    • pp.238-247
    • /
    • 2022
  • Since the skin covers most surfaces of the body, it is susceptible to damage, which can be fatal depending on the degree of injury to the skin because it defends against external attack and protects internal structures. Various types of artificial skin are being studied for transplantation to repair damaged skin, and recently, the production of replaceable skin using three-dimensional (3D) bioprinting technology has also been investigated. In this study, skin tissue was produced using a 3D bioprinter with human skin cell lines and cells extracted from mouse skin, and the printing conditions were optimized. Gelatin was used as a bioink, and fibrinogen and alginate were used for tissue hardening after printing. Printed skin tissue maintained a survival rate of 90% or more when cultured for 14 days. Culture conditions were established using 8 mM calcium chloride treatment and the skin tissue was exposed to air to optimize epidermal cell differentiation. The skin tissue was cultured for 14 days after differentiation induction by this optimized culture method, and immunofluorescent staining was performed using epidermal cell differentiation markers to investigate whether the epidermal cells had differentiated. After differentiation, loricrin, which is normally found in terminally differentiated epidermal cells, was observed in the cells at the tip of the epidermal layer, and cytokeratin 14 was expressed in the lower cells of the epidermis layer. Collectively, this study may provide optimized conditions for bioprinting and keratinization for three-dimensional skin production.

Lupus Heart Disease Modeling with Combination of Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Lupus Patient Serum

  • Narae Park;Yeri Alice Rim;Hyerin Jung;Yoojun Nam;Ji Hyeon Ju
    • International Journal of Stem Cells
    • /
    • v.15 no.3
    • /
    • pp.233-246
    • /
    • 2022
  • Background and Objectives: Systemic lupus erythematosus (SLE) is a chronic autoimmune disease mainly affecting young women of childbearing age. SLE affects the skin, joints, muscles, kidneys, lungs, and heart. Cardiovascular complications are common causes of death in patients with SLE. However, the complexity of the cardiovascular system and the rarity of SLE make it difficult to investigate these morbidities. Patient-derived induced pluripotent stem cells (iPSCs) serve as a novel tool for drug screening and pathophysiological studies in the absence of patient samples. Methods and Results: We differentiated CMs from HC- and SLE-iPSCs using 2D culture platforms. SLE-CMs showed decreased proliferation and increased levels of fibrosis and hypertrophy marker expression; however, HC-and SLE-monolayer CMs reacted differently to SLE serum treatment. HC-iPSCs were also differentiated into CMs using 3D spheroid culture and anti-Ro autoantibody was treated along with SLE serum. 3D-HC-CMs generated more mature CMs compared to the CMs generated using 2D culture. The treatment of anti-Ro autoantibody rapidly increased the gene expression of fibrosis, hypertrophy, and apoptosis markers, and altered the calcium signaling in the CMs. Conclusions: iPSC derived cardiomyocytes with patient-derived serum, and anti-Ro antibody treatment could serve in effective autoimmune disease modeling including SLE. We believe that the present study might briefly provide possibilities on the application of a combination of patient-derived materials and iPSCs in disease modeling of autoimmune diseases.

Establishment of a Simple and Rapid Method for Quantitative Determination of -Aminobutyric Acid Using 1H NMR and Production of γ-Aminobutyric Acid in Cell Suspension Cultures of Coriandrum sativum L. (1H NMR에 의한 γ-Aminobutyric Acid의 간단하고 신속한 정량분석법 확립과 고수 (Coriandrum sativum L.) 현탁배양세포로부터 γ-Aminobutyric Acid의 생산)

  • Kim, Suk-Weon;Liu, Jang-R.
    • Journal of Plant Biotechnology
    • /
    • v.34 no.3
    • /
    • pp.207-212
    • /
    • 2007
  • This study describes a simple and rapid method for quantitative determination of $\gamma$-aminobutyric acid (GABA) using $^1H$ NMR spectroscopy from whole cell extracts of plant suspension cultures. When 9 cell lines derived from 8 species of higher plants maintained in liquid Marashige and Skoog (MS) medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) were subjected to $^1H$ NMR, a cell line of Coriandrum sativum L. exhibited the highest level of GABA. The level reached up to 16.9 mg/dry wt when cells were cultured in MS medium supplemented with 0.5 mg/L 2,4-D after 3 weeks of incubation. The method for quantitative determination of GABA using $^1H$ NMR established in this study could be applied to high-throughput screening of various plant resources for GABA production and the cell suspension culture system of C. sativum could be further developed for commercial production of GABA.

High Cell Density Culture of Bifidobacterium longum by Cross-flow Filtration (Cross-flow filtration에 의한 Bifidobacterium longum의 고농도 배양)

  • Lee, Myong-Suk;Park, Yun-Hee
    • Applied Biological Chemistry
    • /
    • v.40 no.1
    • /
    • pp.18-22
    • /
    • 1997
  • The conditions for production of high cell density of Bifidobacterium longum were investigated and the cross-flow filtration system was used to remove the inhibitory metabolites, lactic acid and acetic acid. The maximum cell growth was observed with glucose as carbon source at the concentration of 50 g/l at $37^{\circ}C$ with the initial pH 6.5. When B. longum was cultured in a cross-flow filtration system, the maximum cell growth was observed at a dilution rate(D) of $0.31\;h^{-1}$ and the dry cell weight was 16.4 g/l($3.5{\times}10^{10}\;cell/ml$), which was about four times higher than that obtained in the batch culture with pH control.

  • PDF

Berberine Production by Cell Suspension Cultures of Cork Tree (Phellodendron amurense Rupr)

  • Choi, Myung-Suk;Shin, Dong-Ill;Park, Young-Goo
    • Korean Journal of Pharmacognosy
    • /
    • v.27 no.1
    • /
    • pp.32-36
    • /
    • 1996
  • Various culture conditions for cell growth and berberine production in cork tree (Phellodendron amurense Rupr.) were investigated. Callus was induced from cambium tissue of cork tree, and cultured on LS liquid medium supplemented with 0.5 mg/1 2,4-D, 0.1mg/1 BA, and 3% sucrose. Several factors enhancing berberine production and cell growth in cork tree cell cultures were found. Some of them enhanced both cell growth and berberine production, but others resulted in a decoupling of cell growth and berberine production with significant in the specific levels. High level of nitrate (80mM), high level of phosphate (8.98mM), and sucrose (7%), 1.0mg/l IAA were effective in berberine production, whereas low level of nitrate (40mM), and phosphate (2.25mM), and high level of sucrose (7%) in the medium were effective in cell growth. Two stage culture(first stage for cell growth, and second stage for berberine production) increased berberine production almost twice (5.06mg/g dry weight) as much as single stage cultures in berberine production.

  • PDF

Stem cell attached 3-dimentional printed polycarprolactone scaffold (줄기세포 탑재 3차원 프린팅 polycarprolactone 스캐폴드)

  • Hong, Gyusik;Cho, Jeong Hwan;Yun, Seokhwan;Choi, Eunjeong;An, Seongmin;Kim, Jung Seok;Lee, Jae Sam;Shim, Jin-hyung;Jin, Songwan;Yun, Won-Soo
    • Journal of the Korea Academia-Industrial cooperation Society
    • /
    • v.20 no.8
    • /
    • pp.618-626
    • /
    • 2019
  • Stem cell therapy is not expected to bestow any therapeutic benefit because of the low engraftment rates after transplantation.Various cell-carrying scaffolds have been developed in order to overcome this problem. When the scaffold is formed by 3-dimensional (3D) printing, it is possible to create various shapes of scaffolds for specific regions of injury. At the same time, scaffolds provide stem cells as therapeutic-agents and mechanically support an injured region. PCL is not only cost effective, but it is also a widely used material for 3D printing. Therefore, rapid and economical technology development can be achieved when PCL is printed and used as a cell carrier. Yet PCL materials do not perform well as cell carriers, and only a few cells survive on the PCL surface. In this study, we tried to determine the conditions that maximize the cell-loading capacity on the PCL surface to overcome this issue. By applying a plasma treated condition and then collagen coating known to improve the cell loading capacity, it was confirmed that the 3% collagen coating after plasma treatment showed the best cell engraftment capacity during 72 hours after cell loading. By applying the spheroid cell culture method and scaffold structure change, which can affect the cell loading ability, the spheroid cell culture methods vastly improved cell engraftment, and the scaffold structure did not affect the cell engraftment properties. We will conduct further experiments using PCL material as a cell carrier and as based the excellent results of this study.

Chromosome Variation in Suspension Cells Derived from Cultured Immature Embryo of Triticum spp. (밀(Triticum spp.)의 미성숙배로부터의 유도한 현탁 배양세포에서의 염색체 변이)

  • 방재욱
    • Journal of Plant Biology
    • /
    • v.33 no.3
    • /
    • pp.189-196
    • /
    • 1990
  • Suspension cell lines have been newly established from the calli derived from the immuature embryo culture of hexapolid (Triticum aestivum var. sicco), tetrapolid (T. durum) and diploid (T. tauchii or Aegilops squarrosa) wheat species. The chromosomal variation in suspension cultured cell lines was examined and old cell line, C82d, established from T. aestivum var. copain was also used. New method using 1-bromonaphthalene for metaphase rapping of suspension cells was developed. Variation in chromosome number was observed among all the suspension lines. Cells with doubled chromosome number and deleted chromosome were also observed. Extensive structural changes in chromosome were found in C82d line. Chromosome aberrations showed loss of chromosome arms and chromosome segment. The mean chromosome number in suspension cells of T. aestivum var. sicco was 40, in C82d line 33, in T. durum 28 and in T. tauchii 14. The stability of chromosome in suspension cells of diploid and tetrapolid wheats was higher than that of hexaploid wheat.

  • PDF