• Development and Reproduction
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• v.11 no.3
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• pp.205-217
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• 2007

As an initial step toward better understanding of the molecular mechanism of estrogen-dependent growth in myoma, an optimal primary cell culture condition has been developed and examined by this study. Myoma and myometrium were cultured by two different methods. Culture stability and $E_2$-responsiveness in stable culture were studied. The culture of digested tissue pieces(Method 2) was found to be a stable culture method for the myoma and myometrium showing a favorable response to estrogen. mRNA expression of PR, IGF-1 and IGF-1 receptor genes was enhanced by $E_2$. The gene responses to $E_2$ were higher in myoma compared with myometrium. Moreover, these responses were more expressive in tissues than in the surrounding cells in primary culture of normal myometrium and myoma, implying a vital role of cell communication through the extracellular matrix in maintaining the estrogen-responsiveness. The development of an improved cell culture system for myoma provides an in vitro tool to further investigate the basis of the tumor formation.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.21 no.3
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• pp.231-239
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• 1999

In oral and maxillofacial surgery, there are many cases requiring the graft of epidermal tissues such as maxillectomy, and vestibuloplasty. There have been so many challenges for the culture of the epidermal tissue. Observing the ultrastructure of the cultured human oral kertinocytes, we could compare this findings with that of in vivo ones. With that, we could find the differencies and similarities between cultured cells and in vivo ones, and evaluate the clinical applications of cultured tissue. Human gingiva was obtained and the specimen was explanted on 24-well plate. Two types of culture media were used in this culture system. One was for the growth of the keratinocytes (Media I), and the other was for the stratification (Media II). Media I had special ingredients for the epidermal growth. Those were 0.5% dimethyl sulfoxide (DMSO), 30ng/ml of epidermal growth factor (EGF), 30ng/ml of cholera toxin, and $5{\mu}g/ml$ of transferrin. We cultured the oral keratinocytes for 3 weeks, and at that time the cultured keratinocytes were processed to prepare the specimen for the TEM study. The results were as follows.; 1. In the phase contrast micrograph, epidermal outgrowth firstly appeared on the 3rd day after explantation, and the growing keratinocytes were activley mitotic, and had polygonal shape and increased N/C ratio. 2. In the phase contrast micrograph, the outer most cells exhibited areas where broad cytoplasmic processes extended out onto the culture subtratum(fan-like appaearance). 3. In the TEM micrographs, the cultured keratinocytes showed stratification. The cells were in elongated form, and there were no morphologic differencies among the layers usually found in the in vivo gingiva. 4. Most of cellular organelles underwent lysis, and keratohyaline granules were seen. Tonofibrils were dispersed in the cytoplasm. 5. The cells were interconnected by desmosomes, and their frequency of distribution was considered to be lower than that of in vivo keratinocytes. 6. We could conclude the cultured oral keratinocytes exhibited signs of terminal differentiation.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.49 no.3
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• pp.232-236
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• 2004

In order to optimize tissue culture conditions for genetic transformation of orchardgrass (Dactylis glomerata L.), the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of a cultivar, 'Roughrider', as explant tissues. The optimal concentration of 2,4-D for the induction of embryogenic callus from mature seeds was 3 mg/L. Plant regeneration frequency was 36.3% when embryogenic calli were cultured on the regeneration medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Addition of 1 g/L casein hydrolysate and 300 mg/L L-proline improved frequencies of embryogenic callus induction and plant regeneration up to 57.3 and 60.7%, respectively. Supplementation of the media with 10 mga $\textrm{AgNO}_3$ and 40 mg/L cysteine enhanced frequencies of callus induction and plant regeneration. Efficient regeneration system established in this study will be useful for molecular breeding of orchardgrass through genetic transformation.

• Archives of Plastic Surgery
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• v.45 no.6
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• pp.542-549
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• 2018

Background Despite the increasing popularity of prosthetic breast reconstruction, scant data exist on the microbiological profile of drainage fluid from closed-suction drains and the relationship thereof to surgical-site infections (SSIs) in breast reconstruction surgery. This study aimed to determine whether bacteria isolated from drainage fluid were associated with the development of SSIs, and whether the bacterial profile of drainage fluid could be a clinically useful predictor of SSIs. Methods We performed a retrospective chart review of 61 women who underwent tissue expander/implant or direct-to-implant reconstructions. Patient demographics and culture studies of drainage fluid from suction drains collected on postoperative day 7 were evaluated. Results Sixteen patients (26.23%) were culture-positive, and 45 patients (73.77%) were culture-negative. The most frequently isolated bacteria were coagulase-negative staphylococci, followed by Staphylococcus aureus. SSIs were diagnosed in seven patients and were mostly resolved by systemic antibiotics; however, the tissue expander or implant was explanted in two patients. Positive culture of drainage fluid from closed-suction drains was significantly associated with the development of SSIs (P<0.05). The positive predictive value was 37.50%, and the negative predictive value was 97.78%. Conclusions To our knowledge, this study is the first to demonstrate a significant association between the microbiological profile of drainage fluid from closed-suction drains and the development of SSIs in patients with prosthetic breast reconstructions. The high negative predictive value suggests that microbial testing of drainage fluid from closed-suction drains may have clinical utility. Further prospective studies with larger sample sizes are required to confirm our findings.

• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.695-703
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• 2006

The dissolved oxygen level of any cell culture environment has a critical effect on cellular metabolism. Specifically, hypoxia condition decreases cell viability and recombinant protein productivity. In this work, to develop CHO cells producing recombinant protein with high productivity, mammalian expression vectors containing a human tissue-type plasminogen activator (t-PA) gene with hypoxia response element (HRE) were constructed and stably transfected into CHO cells. CHO/2HRE-t-PA cells produced 2-folds higher recombinant t-PA production than CHO/t-PA cells in a $Ba^{2+}-alginate$ immobilized culture, and 16.8-folds in a repeated batch culture. In a non-aerated batch culture of suspension-adapted cells, t-PA productivity of CHO/2HRE/t-PA cells was 4.2-folds higher than that of CHO/t-PA cells. Our results indicate that HRE is a useful tool for the enhancement of protein productivity in mammalian cell cultures.

• Korean Journal of Plant Tissue Culture
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• v.21 no.3
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• pp.145-150
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• 1994

This study was conducted to determine the effects of 3 cytokinins (BA,2iP and kinetin) and their concentrations (0, 0.1, 0.5, 1.0, and 2.0 mg/L) on multiple shoot production of Citrus spp. 'Sambokam' and 'Byungkyool' by nodal culture. Nodal explants were obtained from in vitro germinated seedlings of both cultivars. 'Sambokam' produced more multiple shoots than did 'Byungkyool' by nodal culture. Among the 3 cytokinins tested in this study BA supplemented in semi-solid MS basal medium was the most effective stimulator for multiple shoot production, and an optimal concentration was determined to be 1.0 mg/L. Shoot elongation and root formation were inhibited by increasing cytokinin concentration, regardless of cytokinin types. BA at 1.0 mg/L produced the most multiple shoots and the highest number of leaves in 'Sambokam', whereas any cytokinin and concentration studied in this experiment did not affect any scored variables such as shoot and leaf numbers, etc. in 'Byungkyool'.

• Korean Journal of Plant Tissue Culture
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• v.27 no.6
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• pp.429-434
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• 2000

Isolated protoplasts from ginseng (Panax ginseng C. A. Meyer) callus tissue were cultured in modified MS media supplemented with various concentrations of dimethylsulfoxide (DMSO). The cell wall regeneration rate and cell division efficiency of the protoplasts were increased significantly by 1% DMSO treatment. However, there was no difference in the viability of protoplasts between the DMSO treatment and non-treatment. Transmission electron microscopy revealed that the microtubules were oriented in parallel manner to the plasmalemma after 3 days of culture in medium with 1% DMSO. Further, interconnected cellulose microfibrils were observed on the outer surface of the 3-day-cultured protoplasts by scanning electron microscopy These structures shown by electron microscopy were not observed in protoplasts cultured on DMSO-free media. This studies indicates that DMSO supplemented in culture media seemed to stimulate the cell wall regeneration and cell divisions of protoplasts by forming microtubule organizing centers (MTOC).

• Korean Journal of Plant Tissue Culture
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• v.26 no.4
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• pp.271-274
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• 1999

This study was undertaken to develop an efficient propagation technique for mature Betula davurica. Using aseptic materials taken from in vitro culture, the effects of media and plant growth regulators on shoot proliferation and rooting were investigated. DKW medium turned out to be the best in shoot proliferation among the media tested. Whereas axillary buds were better culture material than apical buds in proliferation of shoots, apical buds were slightly better than axillary buds on shoot elongation. Neither 1 /2 MS nor WPM medium seemed to be suitable for shoot multiplication or elongation. When the explants were cultured on 1/2 MS medium, shoot elongation was retarded by forming big callus at the base. In the case of WPM, shoots could be formed normally, but they exhibited slow growing. NAA was so effective on in vitro rooting that more than 80% rooting could be achieved on half-strength DKW medium supplemented with 1.0 mg/L NAA after 4 weeks in cultures. Ex vitro rooting using elongated shoot was also applicable to rooting and acclimatization. Rooted plantlets were successfully acclimatized in an artificial soil mixture and grew normally. The results demonstrate that efficient mass propagation of mature B. davurica can be done through tissue culture.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.28 no.2
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• pp.114-125
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• 2002

This study was designed to evaluate the influence of fibroblasts or connective tissue from mouse oral mucosa on differentiation of neonatal mouse calvaria-derived osteoblasts and mineralization of bone nodules. Primary cell cultures from mouse calvarial osteoblasts and 2-4 passaged fibroblasts from oral mucosa were co-cultured in monolayer cultures, devided into 6 experimental group according to cell density or cell confluency. Osteoblasts were also co-cultured with fibroblasts in $Transwell^{(R)}$ culture plate with different co-cultured period according to osteoblast differentiation. The alkaline phosphatase activity were measured in monolayer cultures and cultures using $Transwell^{(R)}$. The mineralized bone nodules were presented by Von Kossa staining and density of mineralized nodules was measured by image analysis. The connective tissues with or without osteoblast seeding were cultured and examined histologically by Von Kossa and Trichrome Goldner staining. The results were as follows; 1. Prolonged maturation of matrix and delayed mineralization of bone nodules were resulted in monolayer cultures. 2. Co-culture of fibroblast with osteoblast using $Transwell^{(R)}$ during osteoblast proliferation stage stimulated proliferation of osteoblasts and increased alkaline phosphatase activity and mineralization of bone nodules. 3. Co-culture of fibroblast with osteoblast using $Transwell^{(R)}$ during matrix mineralization stage decreased and delayed mineralization of bone nodules. 4. In vitro cultured connective tissue with osteoblast seeding resulted in proliferation of osteoblasts and matrix formation with mineralization.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.416-418
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• 2009

The hepatoprotective effect of Cordyceps militaris culture broth was determined using HaM/ICR strain mice. Compared to control, the intra-peritoneal injection of benzo($\alpha$)pyrene (B($\alpha$)P) remarkably increased the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the serum and the level of lipid peroxide (LPO) in liver tissue, which mean the liver was damaged by B($\alpha$)P. However, compared to B($\alpha$)P, oral administration of C. militaris culture broth showed decrement of AST, ALT, and LPO activities and increment GST activity and GSH level in liver tissue. These suggest that C. militaris culture broth recovered hepatic damage induced by B($\alpha$)P.