• Microbiology and Biotechnology Letters
• /
• v.48 no.2
• /
• pp.205-214
• /
• 2020

Cyanobacteria are microorganisms which have important roles in the nitrogen cycle due to their ability to fix nitrogen in water and soil ecosystems. They also produce valuable materials that may be used in various industries. However, some species of cyanobacteria may limit the use of water resources by causing harmful algal blooms in water ecosystems. Many culture collection depositories provide cyanobacterial strains for research, but their systematic preservation is not well-developed in Korea. In this study, we developed a method for the cryopreservation of the cyanobacteria Trichormus variabilis (syn. Anabaena variabilis), using alginate microcapsules. Two approaches were used for the experiments and their outputs were compared. One of the methods involved the cryopreservation of cells using only a cryoprotectant and the other used the cryoprotectant within microcapsules. After cryopreservation for 35 days, cells preserved with both methods were successfully regenerated from the initial 1.0 × 10⁵ cells/ml to a final concentration of 6.7 × 10⁶ cells/ml and 1.1 × 10⁷ cells/ml. Irregular T. variabilis shapes were found after 14 days of regeneration. T. variabilis internal structures were observed by transmission electron microscopy (TEM), revealing that lipid droplets were reduced after cryopreservation. The expression of the mreB gene, known to be related to cell morphology, was downregulated (54.7%) after cryopreservation. Cryopreservation using cryoprotectant alone or with microcapsules is expected to be applicable to other filamentous cyanobacteria in the future.

• Korean Journal of Clinical Laboratory Science
• /
• v.51 no.3
• /
• pp.360-369
• /
• 2019

Cadherins are essential transmembrane proteins that promote cell-cell adhesion and maintain the corpus luteum structure in the ovary. This study examined the influence of prostaglandin F2 alpha ($PGF2{\alpha}$) on E-cadherin, N-cadherin, and adhesion in luteal theca cells (LTCs). The luteal cells were isolated from the mid-phase corpus luteum, and the LTCs were cultured separately from the luteal heterogeneous cells according to the morphology of the mesenchymal cells and to determine if steroidogenic and endothelial cells of LTCs, 3beta-hydroxysteroid dehydrogenase ($3{\beta}$-HSD), and vascular endothelial growth factor receptor 2 (VEGFR2) mRNA were used. The LTCs were then incubated in the culture medium supplemented with 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ for 24 h, and the E-cadherin and N-cadherin proteins in the LTCs were detected by confocal laser scanning microscopy. The results revealed $3{\beta}$-HSD mRNA expression in the LTC but no VEGF2R mRNA expression. The E-cadherin and N-cadherin proteins of the LTCs were damaged in the 0.01, 0.1, and 1.0 mM $PGF2{\alpha}$ treatment groups, and the expression of the N-cadherin protein was reduced significantly in 0.01 mM $PGF2{\alpha}$ compared to the 0 mM $PGF2{\alpha}$ treatment groups (P<0.05). In addition, the number of attached LTCs were significantly lower in the 0.01 mM $PGF2{\alpha}$ treatment group than in the 0 mM $PGF2{\alpha}$ treatment group (P<0.05). In conclusion, $PGF2{\alpha}$ affected the disruption of cadherin proteins and cell adhesion in LTCs. These results may help better understand the cadherin and adhesion mechanism during corpus luteum regression in the ovary.

• Journal of Life Science
• /
• v.29 no.11
• /
• pp.1227-1234
• /
• 2019

Prunus mume, also known as maesil, is a popular fruit consumed in East Asia (Korea, Japan, and China). It contains high amounts of organic acids, minerals, and polyphenols and has been used as a medication for fever, vomiting, and detoxification. In this study, the anti-proliferative and apoptotic effects of solvent fractions from maesil were evaluated using sulforhodamine B (SRB) assays, morphological evaluations, Hoechst 33258 staining, and western blotting. Addition of the maesil methanol fraction (MMF) and the maesil butanol fraction (MBF) significantly and dose-dependently decreased the cell viability of HT-29 human colon cancer cells. Colony-forming assays confirmed that the MMF and MBF treatments decreased colony numbers when compared with untreated control cells. Treatment of HT-29 cells with MMF and MBF caused a distortion of the cell morphology to a shrunken cell mass. Treatment with MMF and MBF also dose-dependently increased nuclear condensation and the formation of apoptotic bodies in HT-29 cells. Treatment with MMF and MBF significantly and dosedependently increased the expression of Bax (a pro-apoptotic protein), caspase-3, and poly ADP-ribose polymerase (PARP) and decreased the expression of Bcl-2 (an anti-apoptotic protein). MMF significantly and dose-dependently inhibited cell proliferation induced by bisphenol A, an environmental hormone. Therefore, MMF may have potential use as a functional food and as a possible therapeutic agent for the prevention of colon cancer.

• Korean Journal of Poultry Science
• /
• v.40 no.4
• /
• pp.299-304
• /
• 2013

Tracheal epithelial cells (TECs) are an important tool for studies of viral respiratory diseases. Primary TECs have been cultured from human, mouse and hamster. It is also necessary to diagnose viral respiratory disease and reveal infection mechanisms in chicken. In this study, we isolated tracheal epithelial layers from tracheal of 20-day-old chicks and cultured primary TECs from the isolated layers. Ciliated cells which were a typical morphology of TECs were observed in cultured primary TECs and maintained until cell passage 5 (15 to 20 days). When we analyzed expression patterns of epithelial marker genes (retinoic acid responder, FGF-binding protein, virus activating protease (VAP) in TECs compared to immortalized chicken embryonic fibroblast cell line (DF-1), all the marker genes are highly expressed in TECs than in DF-1. When TECs were cultured with 0.1 and 1 MOI of ND virus (rNDV-GFP strain) to test the susceptibility of TECs for ND virus, 12.6% and 48.2% of the incubated TECs were infected respectively. In addition, when DF-1 was incubated with 1 MOI of ND virus, the virus infection rate of DF-1 was three times lower than the virus infection rate of TECs. These data could contribute to study infection mechanisms of viral respiratory diseases and control them in chicken.

• The Journal of Engineering Research
• /
• v.3 no.1
• /
• pp.189-197
• /
• 1998

Hyeroxyapatite(HAp) which has good biocompatibility was made by Wet Chemical Process. The surface of Ti-6Al-4V, coated with HAp by lon Beam Assisted Deposition (IBAD), was treated with 5ppm, 10ppm, 20ppm, and 100ppm of $AgNO_3$ solution. In this Ag impregnation process, $Ca^{2+}$ of HAp was substituted with $Ag^+$ of $AgNO_3$. In this study, the antimicrobial effect and biocompatibility of Ti-6Al-4V alloy which was coated with Ag-HAp were examined. The antimicrobial test was carried out with two kinds of bacteria(P. Aeruginosa, S. Epidermidis), which are highly infectious in a transplanting operation of implant materials. As a result of the test, it was observed that Ti-6Al-4V alloy which was treated by 20ppm of $AgNO_3$ solution has good biocompatibility. In order to observe the antimicrobial mechanism of $Ag^+$, E. coli which is the most common bacterium was treated by Ag-HAp. Then cell morphology of E. coli was observed by the transmission electron microscope(TEM). The destruction of cell wall and cytoplasm of E. coil were observed. A black spot appeared in the cytoplasm was analyzed by energy dispersive analysis X-ray (EDAX) and it showed a small amount of $Ag^+$. Thus, it was proved that $Ag^+$ destroys bacteria effectively and Ti-6Al-4V alloy which was impregnate with Ag ion show antimicrobial effect on infection bacteria.

• Development and Reproduction
• /
• v.11 no.3
• /
• pp.235-243
• /
• 2007

Human umbilical cord blood(HUCB) contains a rich source of hematopoietic stem cells, mesenchymal stem cells and endothelial cell precursors. Mesenchymal stem cells(MSCs) in HUCB are multipotent stem cells, differ from hematopoietic stem cells and can be differentiated into neural cells. We studied on transdifferentiation-promoting conditions in neural cells and cholinergic neuron induction of HUCB-derived MSCs. Neural differentiation was induced by addingdimethyl sulphoxide(DMSO) and butylated hydroxyanisole(BHA) in Dulbeco's Modified Essential Medium(DMEM) and fetal bovine serum(FBS). Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor(bFGF), retinoic acid(RA) and sonic hedgehog(Shh). MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including $\beta$-tubulin III, GFAP and MBP, was markedly elevated during this acute differentiation. The differentiation rate was about $32.3{\pm}2.9%$ for $\beta$-tubulin III-positive cells, $11.0{\pm}0.9%$ for GFAP, and $9.4{\pm}1.0%$ for Gal-C. HUCB-MSCs treated combinatorially with bFGF, RA and Shh were differentiated into cholinergic neurons. After cholinergic neuronal differentiation, the $\beta$-tubulin III-positive cell population of total cells was $31.3{\pm}3.2%$ and of differentiated neuronal population, $70.0{\pm}7.8%$ was ChAT-positive showing 3 folds higher in cholinergic population than neural induction. Conclusively, HUCB-derived MSCs can be differentiated into neural and cholinergic neurons and these findings suggest that HUCB are alternative cell source of treatment for neurodegenerative diseases such as Alzheimer's disease.

• Applied Microscopy
• /
• v.40 no.2
• /
• pp.73-80
• /
• 2010

Liver regeneration is a result of highly coordinated proliferation of hepatocytes and nonparenchymal liver cells. Partial hepatectomy (PH) is the most often used stimulus to study liver regeneration because, compared with other methods that use hepatic toxins, it is not associated with the tissue injury and inflammation, and the initiation of the regenerative stimulus is precisely defined. Granulocyte macrophage-colony stimulating factor (GM-CSF), which is a cytokine able to regulate the proliferation and differentiation of epithelial cells, was first identified as the most potent mitogen for bone marrow. Particularly, rrhGM-CSF, which is highly glycosylated and sustained longer than any other types of GM-CSF in the blood circulation, was specifically produced from rice cell culture. In this experiment, effects of rrhGM-CSF administration were evaluated in the regenerating liver after 78% PH of rats. Morphological changes induced by PH were characterized by destroyed hepatocyte plate around the central vein and enlarged nuclear cytoplasmic ratio and increased hepatocytes with two nuclei. And then, proliferation of liver cells (parenchymal and nonparenchymal) and rearrangement of plates and lobules seemed to be carried out during liver regeneration. These alterations in the experimental group preceded those of the control. Since proliferating cell nuclear antigen (PCNA) is known to be a nuclear protein maximally elevated in the S phase of proliferating cells, the protein was used as a marker of liver regeneration after PH in rats. PCNA levels by western blot analysis and immunohistology were compared between the two groups. PCNA protein expression of two groups at 12 hr and 24 hr after injury showed similar pattern. The protein expression showed the peak at 3 days in both groups, however, the protein level of the experimental group was higher than that of the control. On immunohistochemical observations, the reaction product of PCNA was localized at the nuclei of proliferating cells and the positive reaction in experimental group at 3 days was clearly stronger than that in control group. The results by Western blotting and immunohistology for PCNA showed similar pattern in terms of the protein levels. In conclusion, rrhGM-CSF administration during liver regeneration after 78% PH accelerated breakdown and restoration of the hepatic plate and expression of PCNA. These results suggest that rrhGM-CSF might play an important role during liver regeneration in rats.

• Weed & Turfgrass Science
• /
• v.3 no.3
• /
• pp.196-205
• /
• 2014

In this study, to obtain basic data for searching potential resources as new natural fibers, we investigated morphological and classificatory characteristics of 21 weed seed fibers. According to classification keys in this study, the collected weed seed fibers could be classified into total 13 types, showing their diversity. Seven species among them belonged to BOT3 type. Two species belonged to B2N0 and DOS3 type, respectively. Many of weed seed fibers had not branched. However, three species had various branched fibers at one main fibers on the seed. Three species had various branched fibers at several main fibers on the seed. Eight species had a smooth fiber surface but 13 species had a weakly or significantly developed-corniculum on the fiber surface. In the fiber cell shape, fiber cells of eight weed species were composed of one long cell without septum. But others had a fiber cell shape composed of a bunch of several long cells. Based on the easiness of harvesting, productivity of fibers, and morphological characteristics of seed fiber, it seemed that five seed fibers (TYPLA, METJA, HEMLY, IMPCK, and EREHI) should be additionally investigated if they are practically applicable as renewable resources for new natural fibers.

• Journal of Aquaculture
• /
• v.21 no.4
• /
• pp.331-338
• /
• 2008

Monthly changes in the gonadosomatic index (GSI) and hepatosomatic index (HSI) of wild river puffer Takifugu obscurus, and water quality environment in spawning area during breeding season were investigated from March 1995 to February 1996. Monthly changes in GSI and HSI of T. obscurus, that was cultured in low salinity, were calculated. The external morphology of the gonads, germ cell differentiation during gametogenesis and the reproductive cycle with the gonad developmental phases were investigated by histological analysis. The optimum water quality environment in Ganggyung, Choongcheongnam-do, where is spawning ground of wild T. obscurus, was $15-20^{\circ}C$ (water temperature) and 0 psu (salinity). Monthly changes in the GSI in females and males reached a maximum in May, and then rapidly decreased. Therefore, it is assumed that in the natural condition the spawning period of wild T. obscurus is May to June. In females and males, it showed a negative correlationship between the GSI and HSI. The external morphology of the gonads in female and male T. obscurus, that was cultured in low salinity, is composed of a pair of saccular structure. Based on monthly changes in the GSI, it is assumed that in female T. obscurus, that was cultured in low salinity, spawn from March through May. Therefore, it showed a negative correlationship between changes in the GSI and HSI. On the whole, in females and males, it showed a similar pattern between wild and cultured T. obscurus. The reproductive cycle with the gonad developmental phases can be classified into successive five stages in females: the early growing stage, late growing stage, mature stage, ripe and spent stage, and recovery and resting stage. In males, that can be divided into successive four stages: the growing stage, mature stage, ripe and spent stage, and recovery and resting stage. In case of wild T. obscurus, the spawning period has once a year, however, those cultured in the high water temperature ($20-27^{\circ}C$) - low salinity (under 3.3 psu) condition have reproductive characteristics having possibilities of discharge of eggs and sperms year-round as a multiple spawner.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.24 no.1
• /
• pp.1-17
• /
• 1997

The purpose of this study was to investigate the aspects of proliferation and bone nodule formation of osteogenic precursor cells. To determine the effects of ascorbic acid and dexamethasone upon capacity of osteoblast proliferation and bone nodule formation, cells were maintained in the presence of one or some of these additives for up to 30 days. Group I culture was maintained in standard medium(DMEM plus 10% plus antibiotics), group II was maintained in supplemented medium containing dexamethasone, group III was maintained in supplemented medium containing ascorbic acid and sodium-${\beta}$-glycerophosphate, and group IV was maintained in supplemented containing ascorbic acid, sodium-${\beta}$-glycerophosphate and dexamethasone. Morphology of bone nodules was observed with light microscope and electron microscope. The results were as follows: ${\bullet}$ Proliferation capacity of osteoblasts was not affected by single use of dexamethasone, but it was chiefly affected by ascorbic acid. ${\bullet}$ Cellular morphology was fibroblastic appearance initially, but, it was gradually changed to polygonal shape accompanied by confluency stage. ${\bullet}$ Pluripotent mesenchymal cells existed during primary culture, they were differentiated to adipocyte, chondrocyte, osteocyte according to culture condition. ${\bullet}$ Dexamethasone increased bone nodule formation under the condition that the culture was maintained with supplemented medium ascorbic acid and sodium-${\beta}$-glycerophosphate. ${\bullet}$ when the cultures were stained with alizarin red, the group supplemented with dexamethasone, ascorbic acid and sodium-${\beta}$-glycerophosphate showed the marked increase of bone nodule formation, but the group supplemented with ascorbic acid and sodium-${\beta}$-glycerophosphate revealed only small amounts of bone nodules. And the groups cultured without ascorbic acid showed no observed any of bone-like mass independent of dexamethasone addition.