Background: Cytokine-mediated ex vivo expansion has been proposed as a means of increasing the number of cord blood (CB) hematopoietic stem cells for transplantation. As well as stem cell number, stromal cells are necessary for functional maturation of hematopoiesis. The purpose of this study was to analyze the development of stromal cells during ex vivo expansion of CB $CD34^+$ cells. Methods : $CD34^+$ cells were purified from CB by magnetic bead selection. The levels of of interleukin-3, interleukin-$1{\beta}$, interleukin-6, granulocyte macrophagecolony stimulating factor and tumor necrosis factor-${\alpha}$ were measured in culture supernatants on 0, 1, 2, and 3 weeks, using ELISA techniques. CB $CD34^+$ cells were expanded in Iscoves modified Dulbeccos medium in the presence of several cytokines. The expression of E-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, platelet/endothelial cell adhesion molecule-1, von Willebrand factor, vimentin, and CD14 in newly developed stromal cells was examined by immunocytochemical method. Relevant extracellular matrix (ECM) proteins and proper cytokines were also assayed for the most suitable condition for expansion of stromal cells. Results: Several cytokines were found to have been produced by CB $CD34^+$ cells as well as bone marrow-derived $CD34^+$ cells. During ex vivo expansion of CB $CD34^+$ cells, stromal cells appeared in the culture by day 4 and expanded over the following 7-10 days before being confluent by day 2 1. These cells expressed surface markers characteristic of cells of endothelial lineage. Furthermore, these stroaml cells also expanded effectively when treated with thrombopoietin+flt-3 ligand+stem cell factor+leukemia inhibitory factor or 0.1% poly-L-lysine-coated wells. Conclusion: Stromal cells were developed during ex vivo expansion of CB $CD34^+$ cells and that this development could be enhanced further by treating the stromal cells with cytokines or ECM.
Choi, Gyeong Lee;Rhee, Han Cheol;Yeo, Kyung Hwan;Lee, Seong Chan;Kang, Nam Jun;Choi, Hyo Gil
Journal of Bio-Environment Control
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v.26
no.1
/
pp.1-6
/
2017
The paprika has emerged as one of the highest-income crops by increase in dimestic and export demand in the greenhouse crops. Nevertheless, there is no standard for fertigation in soil, because general culture system is soilless culture. This study was conducted to establish the optimum nitrogen and potassium application level for paprika fertigation. Four different levels of nitrogen and potassium were applied, treatment levels were 0.5, 1.0, 1.5, 2.0 times of pimiento fertilization recommendations based on soil testing. Experiment to instigate the optimum amounts of nitrogen and potassium were carried out in 2012 and 2013, respectively13. Nitrogen application : stem diameter of 0.5 times was significantly lower than other treatments, but stem length was not affected by nitrogen fiertigation levels. Number of fruit and yield of first fruiting group harvest were not significant difference. but those of the second fruiting group were decreased by increasing nitrogen level beyond 1.0 times treatment and were the lowest in 0.5 times treatment. Overall, the optimum level of nitrogen for fertigation was judged 1.0 times of pimiento fertilization recommendations based on soil testing. Potassium application : Growth was no signigicant trend except stem length. Number of locule, fresh thickness and sugar content were not significant difference. Number of fruit and yield were not significant difference at the first and second fruiting group harvest. But those were significant difference at third fruiting group harvest, maximum yield was obtained by 1.5 times fertigation level. The optimum level of potassium for fertigation was judged 1.5 times of pimiento fertilization recommendations based on soil testing.
This experiment was carried out to study the effects of nitrogen concentration of cultural solution, Rhizobium inoculation, and planting density on the growth and yield of soybean cultivars, Hwanggeumkong, Jangbaegkong, Paldalkong, Clark, and non-nodulation isoline of Clark. Rhizobium inoculation increased the stem length, particularly in Hwanggeumkong, Jangbaegkong, and decreased it significantly in non-nodulation Clark. Stem length was increased by the increase in nitrogen fertilization by the 195ppm level and decreased by the increase in plant population density. Rhizobium inoculation also increased the shoot dry weight, but significantly decreased it in non-nodulation Clark. As nitrogen concentration in the cultural solution increased the shoot dry weight decreased in Jangbaegkong and paldalkong. However, the shoot dry weight was decreased by the increase in plant population density. Rhizobium inoculation and the increase in nitrogen concentration of cultural solution increased the ratio of shoot dry weight to root weight. The Rhizobium inoculation and the increase in nitrogen concentration of cultural solution increased the grain yield per pot in Hwanggeumkong and paldolkong, While non-nodulating Clark showed significant decrease in grain yield. Grain yield per pot was also increased by the increase of plant population density. Grain yield was significantly correlated with shoot dry weight, nodule number, and nitrogen content of the soybean plants. The correlation between nitrogen contents of the soybean plants and stem length, shoot dry weight, and nodulation was significant. The allantoin-N content in stem was also significantly correlated with nodulation.
Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.
Kim, Yun-Hui;Lee, Dong-Soo;Kang, Joo-Hyun;Lee, Yong-Jin;Chung, June-Key;Lee, Myung-Chul
The Korean Journal of Nuclear Medicine
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v.38
no.1
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pp.99-108
/
2004
Purpose: The ability to noninvasively track the migration of neural progenitor cells would have significant clinical and research implications. We generated stably transfected F3 human neural progenitor cells with human sodium/iodide symporter (hNIS) for noninvasively tracking F3. In this study, the expression patterns of hNIS gene in F3-NIS were examined according to the cultured time and the epigenetic modulation. Materials and Methods: F3 human neural stem cells had been obtained from Dr. Seung U. Kim (Ajou University, Suwon, Korea). hNIS and hygromycin resistance gene were linked with IRES (Internal Ribosome Entry Site) under control of CMV promoter. This construct was transfected to F3 with Liposome. To investigate the restoration of hNIS gene expression in F3-NIS, cells were treated with demethylating agent (5-Azacytidine) and Histone deacetylase inhibitor (Trichostatin A: TSA). The expression of hNIS was measured by I-125 uptake assay and RT-PCR analysis. Results: The iodide uptake of the F3-NIS was higher 12.86 times than F3 cell line. According to the cell passage number, hNIS expression in F3-NIS gradually diminished. After treatment of 5-Azacytidine and TSA with serial doses (up to $20{\mu}M$, up to 62.5nM, respectively) for 24 hours, I-125 uptake and mRNA of hNIS in F3-NIS were increased. Conclusion: These results suggest that hNIS transfected F3 might undergo a change in its biological characters by cell passage. Therefore, the gene ex[ressopm of exogenous gene transferred human stem cell might be affected to the epigenetic modulation such as promoter methylation and Histone deacetylation and to the cell culture conditions.
Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.
'Secret Pink' was released by National Institute of Horticultural & Herbal Science in 2008. The cross was made between '03B1-23' and 'Sei-Monaco in 2005'. After the evaluation of the characteristics under shade culture in summer and retarding culture in spring and consecutive selection from 2006 to 2008, 'Secret Pink' was selected finality. The natural flowering time of 'Secret Pink' was the middle of October, and it is possible to flower all year round by shade and light culture. It has single type flowers with pink petals. The growth of plant was very vigorous and response time was 6.5 weeks. The diameter of flower was 6.7 cm. Number of flowers per stem and petals per flower were 14.5 and 25, respectively. Days to flowering under the short day treatment was about 50 and its vase life was 25.5 days in autumn season.
This study was carried out to investigate the effects of nutrient solution concentration, irrigation frequency on growth, flowering, and cut flower longevity of Dendranthema grandiflorum (Ramat.) Kitamura 'Shuhouno-chikara' hydroponically grown in perlite media. Not only stem length but also mineral contents of each plant organ in nutrient solution culture were much higher than those in soil culture. 1/2S of nutrient solution was good at early stage, but 1S of nutrient solution was better as chrysanthemum growth progressed. Among different concentrations of nutrient solution, mineral contents of each plant part showed no significant difference. 1S treatment of nutrient solution increased the vase life by 3 days than 2S treatment. The growth and mineral contents of each plant organ were great in plants irrigated 8 times a day, because of high moisture contents of medium and high water availability, followed by more frequent nutrient replenishment near roots. There was no correlation between nutrient solution concentration and vase life of cut flower grown in nutrient solution culture. Cut flowers irrigated twice per day had the longer vase life than other treatments (4, 8 times).
We found out the new method of the consortium for the environmental friendly agriculture by 8 kinds of the selected antagonistic rhizobacteria. This research involved composition of mutual complementary consortium by each antagonistic function such as production of antibiotic, siderophore, antifungal cellulase and insoluble phosphate solubilization. The consortium No.11 among composed consortium candidates showed the most pepper growth promoting activity and Phytophthora blight suppression on the in vivo pot test of red-pepper plant. The consortium No. 11 is combination of PGPR Bacillus subtilis AH18 and Bacillus licheniformis K11. B. subtilis AH18 and B. licheniformis K11 both could produce the auxin, antifungal ${\beta}$-glucannase and siderophore. Also, they had mechanism for solubilization of insoluble phosphate. But, B. licheniformis K11 could produce the antibiotic of iturin which was able to inhibit Phytophthora capsici. We confirmed complementary noncompetitive mutualism between B. subtilis AH18 and B. licheniformis K11 of the consortium No.11. The results came out through treatment of two strains co-culture, treatment of individual culture and co-treatment of two individual cultures for the growth and Phytophthora blight suppression of red-pepper. The treatment of two strains co-culture didn't show a synergic effect in comparing sole treatment on the pepper growth promotion and Phytophthora blight suppression. But, when the pots were treated simultaneously with co-treatment of two individual cultures, an synergic effect was seen in the growth promotion of roots, stem, leaves and suppressed Phytophthora blight on red-pepper in vivo pot test.
This study was aimed to proliferate Gentiana axillariflora Leveille which is one of the important medicinal and ornamental plants, by establishment of multiple shoot formation and embryogenesis through tissue culture technique. Callus was formed on MS (Murashige and Skoog) medium supplemented with 2,4-D, CPA, but not formed with BAP. The addition of 2,4-D 2 mg/ l into the medium was effective for callus formation and the rate of callus formation was about 90%. Somatic embryos were obtained on MS medium for two months. When callus was cultured on MS medium with combination treatment of 2,4-D 0.5 mg/ l and BAP 0.5 mg/ l, the number of embryo formed was better than that of other single or combination treatments and the total numbers of embryo a were 18.8 (number of total embryo/number of explants incubated = 753/40) at mean. Callus induction from stem and node explants was increased by addition of TDZ 2 mg/ l in the presence of 2,4-D 2 mg/ l, respectively. The best result about the differentiation of shoots was obtained on MS medium added BAP 2 mg/ l from node culture. Multiple shoots from shoot apex were induced on MS medium containing BAP 1 mg/ I and TDZ 1 mg/ l , BAP 2 mg/ l and TDZ 1 mg/ l. The number of multiple shoots per one explant was above seventy plants. It was the most effective regeneration system for rapid multiplication of Gentiana axillariflora Leveille.
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