• Polymer(Korea)
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• v.29 no.3
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• pp.294-299
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• 2005

Thermo-responsive poly(N-isopropylacrylamide) (PIPAAm) was covalently patterned by masked el electron beam irradiation. Introduction of PIPAAm on tissue culture polystyrene dish was confirmed by ATR-FTIR and ESCA measurements. Hepatocytes were cultured at $37^{circ}C$ on these surfaces. Cells adhered on PIPAAm-grafted domains were detached by reducing culture temperature to $20^{circ}C$. Endothelial cells were then seeded and cultured on the same surfaces. Seeded endothelial cells were selectively attached on hepatocytes detached and PIPAAm-grafted domains and could be co-cultured with hepatocytes on the same culture dishes with clear pattern. This co-culture method enabled long-term co-culture of hepatocytes with endothelial cells.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.77-81
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• 1994

To improve in vitro embryonic cell development, this study was desigend to culture in vitro fertilized early embryos of mouse in two different systems; conditioned medium alone and ampullary cells co-culture. Thirty two of 83 embryos(38.6%) were blocked in the 2 cell stage by co-culture, as compared to forty of 42 embryos(95.2%) in control group for 24hours culture. And all the embryonic cells cultured for conditioned medium alone were blocked for 48 hours culture. Twenty seven of 46 embryos (58.7 %) which overcome culture block in 2 cell stage by cocultured were developed morular and expanded blastocyst, and ninteen of 46 embryos(26.1 %) underwent hatching for 96 hours culture. The cellular fragmented rates for embryo were 26.2% in medium alone; 10 fragmented blastomere were graded mild status and 1 fragmented blastomere in severe status. On the other hand, the fragmented rate for 48 hours co-cultured were 15.7%03/83); 8 fragmented embryos were graded mild status, moderate status in 3 fragmented embryos and severe in 2 fragmented embryos respectively. In conclusion, the co-culture of embryos with ampullary cells is good to improve quality of embryos and overcome of culture block as well as development of cell cleavage.

• Journal of Embryo Transfer
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• v.15 no.2
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• pp.129-136
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• 2000

This study was carried out to investigate the effect of biological factors on the in vitro production(IVP) of bovine oocytes for development of simple culture methods and medium. Oocytes from the slaughterhouse ovaries were matured and fertilized using general protocol and this study was examined if there were necessary to co-culture, media change, media type and embryo density. This results were as follows: 1. The development rate according to co-culture with cumulus cells and non co-culture as drop culture was not significantly different in cleavage (68.9 vs 71.7%), 8-cell stage (41.2 vs 44.1%) and blastocyst stage (12.2 vs 13.8%), respectively (p<0.05) 2. The blastocyst development rates in YS and CRIaa were higher than that in TCM199 (12.4, 10.4$ vs 3.7%), but the cleavage (69.0, 77.8 and 61.0%) and 8-cell stage (31.7, 37.0 and 35.7%) development accoring to YS, TCM199 and CRIaa ws not significantly different, respectively (p<0.05). 3. There was no significantly different in cleavage (62.6, 59.5 and 61.2%), 8-cell(34.7, 37.9 and 34.0%) and blastocyst (9.5, 11.6 and 12.8%) development among medium change time as control, Group I and Group II, respectively (p<0.05). 4. Blastocyst formation of 8-cell stage according to embryo density was not significantly different in 1, 10 and 25 embryos (27.3, 22.5 and 34.0%), respectively (p<0.05). These results indicated that simple culture system could reduce bovine IVP embryos as drop culture as non co-culture system, high density embryo (25 embryos/50 $\mu$1 drop). YS defined medium and no medium change for whole culture period, although other biological factors need to examine in order to produce efficient IVP bovine embryos.

• Journal of Animal Science and Technology
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• v.54 no.2
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• pp.103-109
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• 2012

Present study was performed to investigate the effect of single and co-culture of adipocyte and muscle cell lines on cell differentiation. 3T3-L1 (adipocyte) and L6 (muscle) cell lines were single-cultured on the condition of 10% fetal bovine serum (FBS)/Dulbeco's modified eagle's medium (DMEM) for 48 h followed by culture within 5% FBS/DMEM as a growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without additives in single- or co-culture of the 3T3-L1 and the L6 cells to induce differentiation of both cell types. In co-culture system, the 3T3-L1 and the L6 cells were grown in separated places by being seeded on a $0.4{\mu}m$ insert membrane and on the bottom of 6 well plate, respectively. Cell differentiation was measured using morphological investigation and cytosolic analysis of glycerol-3-phosphate dehydrogenase (GPDH; for 3T3-L1) and creatine kinase (CK; for L6). Based on the GPDH results, the presence of L6 cells did not stimulate 3T3-L1 differentiation showing more differentiation of 3T3-L1 cells in the single-culture compared to the co-culture condition. In contrast, 3T3-L1 cells in the co-culture promoted differentiation of L6 cells. Enzymatic analysis supported this result showing that 3T3-L1 cells showed statistically (P<0.05) higher GPDH activity in the single-culture than the co-culture, whereas CK results of L6 cells were vice versa (P<0.05). Overall, present results may indicate that co-culture system is more reliable and precise technique compared to single-culture. Further studies on several co-culture trials including different media conditions, supplementation of differentiating substances, molecular biological analysis, etc. should be required to obtain practical and fundamental mass data.

• Archives of Pharmacal Research
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• v.27 no.1
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• pp.106-110
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• 2004

The antioxidant activities of Acanthopanax senticosus stems were evaluated in $CCl_4$-intoxicated rats. The n-butanol fraction from the water extract of the stems, when pretreated orally at 200 mg/kg/day for 7 consecutive days in rats, was demonstrated to exhibit significant increases in antioxidant enzyme activities such as hepatic cytosolic superoxide dismutase, catalase and glutathione peroxidase by 30.31, 19.82 and 155%, respectively. The n-butanol fraction whereas showed a significant inhibition of serum GPT activity (65.79% inhibition) elevated with hepatic damage induced by $CCl_4$-intoxication. Eleutheroside B, a lignan component, isolated from the n-butanol fraction was found to cause a moderate free radical scavenging effection DPPH, its scavenging potency as indicated in $IC_{50}$ value, being 58.5$\mu$ M. These results suggested that the stems of A. senticosus possess not only antioxidant but also hepatoprotective activities.

• Journal of Embryo Transfer
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• v.16 no.1
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• pp.41-46
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• 2001

This study was carried out to evaluate the effect of culture methods on development of embryos with each developmental stage after heat shock in bovine oocytes. The results obtained were as fellows. 1. The culture method after heat shock on development of embryos was better drop-culture than co-culture. 2. The medium without amino acids were not effect of heat sock on development of embryos but it was in need of amino acid during formation of blastocyst.

• Reproductive and Developmental Biology
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• v.29 no.4
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• pp.223-227
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• 2005

In vitro maturation of porcine immature cumulus-enclosed oocytes can be enhanced by co-incubation with spermatozoa even before fertilization. The aim of this study was to determine whether the addition of spermatozoa into the culture medium can stimulate the meiosis resumption of porcine cumulus-enclosed oocytes arrested at germinal vesicle (GV). Cumulus-enclosed oocytes (CEOs) were collected from follicles of 3 to 5mm diameter. Porcine CEOs were cultured in tissue culture medium containing various meiosis inhibitors and spermatozoa. Oocytes were examined for evidence of GV and GV breakdown after 24 h culture. After 24 h culture $43.8\%$ of oocytes cultured in only TCM 199 remained at GV stage whereas $56.2\%$ of oocytes were able to resume meiosis. When porcine CEOs were cultured in the medium with meiosis inhibitor such as, dibutyryl cAMP (dbcAMP) and forskolin (Fo), more than $90\%$ of oocytes were not able to resume meiosis. However, co-culture of porcine CEOs with spermatozoa was able to overcome the inhibitory effect of dbcAMP and Fo. Irrespective of the presence of 3-isobutyl-1-methylxanthine (IBMX), no difference was observed in the proportion of oocyte reached germinal vesicle breakdown (GVBD). The present study suggests that dbcAMP and Fo prevent the spontaneous maturation of competent oocyte in culture after isolation from follicles and that mammalian spermatozoa contain a substance(s) that improves meiosis resumption in vitro of porcine cumulus-enclosed oocytes.

• Food Science and Preservation
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• v.14 no.1
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• pp.94-99
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• 2007

To investigate the effects of Pichia anomala SKM-T and Galactomyces geotrichum SJM-59 on Baechu kimchi fermentation, lyophilized yeasts were added to Baechu kimchi and co-cultured at room temperature ($20{\pm}2^{\circ}C$) for 7 days. Desirable pH and acidity levels appeared by 3 days of fermentation in both the control culture and that with added G. geotrichum SJM-59. Furthermore, the culture with G. geotrichum SJM-59 sustained a desirable pH and acidity level until 5 days of co-culture. The pH of the culture with P. anomala SKM-T decreased slowly and was significantly higher than that of control throughout the experimental period. As fermentation time increased, the acidity of the culture with P. anomala SKM-T increased gradually. However, this culture maintained a desirable acidity level throughout the experiment. The number of lactic acid bacteria in the culture with P. anomala SKM-T was higher than in the culture with G. geotrichum SJM-59, or the control culture, throughout the experiment. The highest LA/TM ratio appeared after 3 nays of fermentation in the control culture, and on the 5 day of the yeasts added co-cultures. On sensory evaluation, no differences were detected between control and the culture with G. geotrichum SJM-59 arter 3 days of fermentation. The co-cultures with yeasts received high marks in umami taste. The co-culture with P. anomala SKM-T showed better texture properties than did the control culture. It was considered that fermentation times were delayed by addition of G. geotrichum SJM-59 or P. anomala SKM-T to Baechu kimchi fermentation.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.3
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• pp.146-151
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• 2016

Objective: This study was conducted to compare the effects of static culture, dynamic culture, and the combination of dynamic culture with specialized surfaces involving co-culture on human embryonic development. Embryos cultured using conventional static culture (SC) techniques served as a control group. We compared dynamic culture using micro-vibration culture (MVC) and micro-vibration with co-culture (MCoC), in which autologous cumulus cells were used as a specialized surface. Methods: We conducted a chart review of patients who were treated between January 2011 and November 2014 in order to compare embryonic development rates and pregnancy rates among the groups. Zygotes were cultured in micro-droplets, and embryos were subsequently selected for transfer. Some surplus embryos were cryopreserved, and the others were cultured for blastocyst development. A micro-vibrator was set at the frequency of 42 Hz for duration of 5 seconds per 60 minutes to facilitate embryo development. Results: No significant differences among the groups were present in patient's characteristics. However, the clinical pregnancy rates were significantly higher in the MVC group and the MCoC group than in the SC group. No significant differences were found in the blastocyst development rate between the SC group and the MVC group, but the blastocyst development rate in the MCoC group was significantly higher than in the SC and MVC groups. Conclusion: The clinical pregnancy rate was significantly increased by the application of micro-vibration to the embryonic cultures of poor responders. The blastocyst development rate was significantly increased by the application of MCoC to surplus embryos.

• Biomedical Science Letters
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• v.22 no.1
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• pp.1-8
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• 2016

Several studies have investigated the various effects of dexamethasone (Dex) on the proliferation and differentiation of mesenchymal stem cells (MSCs). Previously, we reported that co-treatment with L-ascorbic acid 2-phosphate and fibroblast growth factor (FGF)-2 maintained differentiation potential in MSCs through expression of hepatocyte growth factor (HGF). In this study, we investigated the effects of co-treatment with FGF-2 and Dex on the proliferation and differentiation potential of MSCs during a 2-month culture period. Co-treatment with FGF-2 and Dex increased approximately a 4.7-fold higher accumulation rate of MSC numbers than that by FGF-2 single treatment during a 2-month culture period. Interestingly, co-treatment with FGF-2 and Dex increased expression of HGF and maintained adipogenic differentiation potential during this culture period. These results suggest that co-treatment with FGF-2 and Dex preserves the proliferation and differentiation potential during long-term culture.