• Reproductive and Developmental Biology
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• v.28 no.2
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• pp.101-105
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• 2004

This study was carried out to evaluate the effects of glucose and sodium phosphate on in vitro development of porcine oocytes matured and fertilized in vitro. When the culture medium was supplemented with various concentrations of glucose, the higher proportions (23 and 26%) of oocytes developed to morular or blastocyst stages were at the concentrations of 2.78 and 5.56 mM than 0 (9%; P<0.05) and 11.12 mM (18%). In experiment to evaluate effect of sodium phosphate during in vitro development of porcine oocytes, a significantly (P<0.05) higher proportions of embryos developed to morular or blastocyst stages was obtained with sodium phosphateof 0.28 (25%) and 0.53 (27％) mM than 0 (15%), 1.05 (19%) and 2.10 (10%) mM. On the other hand, when oocytes were cultured in medium with (0.53 mM) sodium phosphate, the proportions of developed embryos were significantly (P<0.05) higher in medium without (29%) that than with (14%) 5.56 mM glucose. However, a higher proportion of embryos developed to morular or blastocyst stages were obtained in medium with (23%) that than without (8%) glucose (P<0.05). The minimum essential medium (MEM) added to the culture medium were higher regardless of presence of sodium phosphate and glucose on the development of embryos. Although sodium phosphate and glucose could support morular and blastocyst development to a limited extend (10∼24%), significantly higher proportion (36%) at morular or blastocyst stages was obtained by MEM adding in the medium with sodium phosphate and glucose. These results suggest that the early development of in vitro fertilized porcine oocytes can be maintained efficiently by glucose and sodium phosphate when they were cultured in medium with MEM.

• Korean Journal of Microbiology
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• v.42 no.2
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• pp.131-134
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• 2006

The culture conditions for the enhanced mycelial of Clavicorona pyxidata DGUM 29005 were investigated. The optimal temperature and pH for the mycelial growth were $24^{\circ}C$ and 5.0, respectively. It was shown that trehalose was the best supplement of carbon sources in Czapek-Dox medium as a minimal medium for enhanced mycelial growth. In general, inorganic nitrogen sources were better than organic ones for mycelial growth. Calcium nitrate was the best out of the inorganic nitrogen test. The appropriate phosphorous and vitamin were $Na_2HPO_4$ and p-aminobenzoic acid, respectively. After the mycelial of C. pyxidata DGUM 29005 was cultivated at $24^{\circ}C$ for 20 days in MEM broth(pH 5.0), the specific activities of both exomycelial and endomycelial enzymes were determined. Among the exomycelial enzyme assayed, the specific activity of laccase was much higher than those of other enzymes. However, little or no enzyme activities of ${\alpha}$-amylase, chitinase, lipase and pretense were found.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.8-8
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• 2001

Mouse follicles require the addition of gonadotropins (Gns) to complete maturation and ovulation of oocyte and antrum formation of follicles in vitro. However, we tried examination of in vitro growth of mouse pre-antral follicles in medium without Gns and physiological factors. And also, pre-antral follicles were isolated from ovaries by mechanical method. Our present studies were conducted to evaluate on the growth of follicles and intra-follicular oocytes and antrum formation in vitro of mouse pre-antral follicles in two different media. Pre-antral follicles (91-120${\mu}{\textrm}{m}$) were isolated mechanically by fine 30G needles not using enzymes from ovary of 3-6 weeks old female ICR mice. Isolated pre-antral follicles were cultured in 20 ${mu}ell$ droplets of TCM (n=17; follicles: 107.8 $\pm$ 1.58 ${\mu}{\textrm}{m}$; oocytes: 59.9$\pm$1.2 ${\mu}{\textrm}{m}$) or MEM (n=12; follicles: 109.3$\pm$2.53 ${\mu}{\textrm}{m}$; oocytes: 55.4 $\pm$1.6${\mu}{\textrm}{m}$) under mineral oil on the 60mm culture dish. All experimental media was supplemented with 10% FBS but without Gns and/or physiological factors. Pre-antral follicles were individually cultured in drops for 8 days. Antrum formation and growth of pre-antral follicles and intra-follicular oocytes were evaluated using a precalibrated ocular micrometer at $\times$200 magnifications during in vitro culture. Results between different groups were analyzed using combination of Student's t-test and Chi-square, and considered statistically significant when P<0.05. Antrum formation of pre-antral follicles had started in two culture media on day-2. On day-8, antrum formation had occurred in 58.3%(7/12) of pre-antral follicles cultured in MEM, but only in 23.5% (4/17) of those cultured in TCM (P=0.0364). Growth of pre-antral follicles and intra-follicular oocytes were observed on day-4 and -8. On day-4, follicular diameters was similar (P=0.1338) in TCM (119.4$\pm$2.58 ${\mu}{\textrm}{m}$) and MEM (125.4$\pm$4.52 ${\mu}{\textrm}{m}$). However, on day-8, diameters of pre-antral follicles cultured in MEM (168.9$\pm$17.29 ${\mu}{\textrm}{m}$) was significantly (P=0.0248) bigger than that in TCM (126.7$\pm$4.28 ${\mu}{\textrm}{m}$). On day-4 and -8, diameters of intra-follicular oocytes were similar TCM (67.1$\pm$1.3 and 72.4$\pm$0.9${\mu}{\textrm}{m}$) and MEM (65.2$\pm$1.7 and 73.3$\pm$1.5 ${\mu}{\textrm}{m}$), respectively. We can conform that medium not supplemented with Gns and/or physiological factors can be used for in vitro antrum formation and growth of mouse pre-antral follicles and intra-follicular oocytes. In conclusion, MEM supplemented with FBS can be used for growth in vitro of mouse pre-antral follicles isolated mechanically.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.5
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• pp.667-673
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• 2009

Ninety six intact male sheep (12 months old with mean live weight of about 35 kg) were used to assess the effects of restricted feeding on intake, digestion, nitrogen balance and metabolizable energy (ME). The animals were selected from two known Iranian small and large body size breeds: 48 Sangsari (S) and 48 Afshari (A), and were divided into two equal groups: restricted (R) and a control (C). Each group had 48 sheep (24 each breed). The experiment had a duration of 15 and 75 days adaptation and treatment periods, respectively. The animals were individually placed in metabolism cages and fed a diet based on pelleted concentrate mixture consisting of alfalfa, barley grain, cottonseed meal and barley straw. The animals in group C were fed ad libitum, while animals in group R were fed at maintenance level and maintained a relatively constant live weight. During the experiment, the average daily weight gain (ADG) of S and A animals in R group was 0.34 and -0.25 g/d (0.02 and -0.02 $g/kg^{0.75}/d$), respectively. While that of S and A animals in C group was 174.4 and 194.4 g/d (10.16 and 11.48 $g/kg^{0.75}/d$), respectively. Nitrogen (N) was determined by both measured and regression methods. Animals of R group stayed at about zero N balance (0.01 and -0.00 g $N/kg^{0.75}/d$ for S and A animals, respectively). The N retention of animals of both S and A breeds in C group were similar (0.45 and 0.46 g $N/kg^{0.75}/d$, respectively). Digestible organic matter intake (DOMI) and ME requirement for maintenance (MEm) were measured by both constant weight technique and regression method by regressing N balance on DOMI and ME intake on ADG. The measured DOMI during constant weight was 24.61 and 24.27 g $DOMI/kg^{0.75}/d$ and the calculated DOMI from regression equation was 24.24 and 24.22 g $DOMI/kg^{0.75}/d$, for S and A animals, respectively. The measured MEm was 402 and 401 kJ $ME/kg^{0.75}/d$ and the calculated MEm from regression analysis was 398 and 400 kJ $ME/kg^{0.75}/d$ for S and A breeds, respectively. There were no significant differences between both measured and regression techniques. There was no significant difference between S and A breeds for DOMI, N retention, MEm, digestibility and metabolizability values. Digestibility values for OM, GE and CP and metabolizability were significantly (p<0.05) higher in restricted feeding sheep compared with that of sheep fed ad libitum.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.100-103
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• 2004

The culture condition and medium composition for the enhanced mycelial growth of Inonotus mikadoi IMSNU 32058 were investigated. The optimal temperature and pH for the mycelial growth were $27^{\circ}C$ and pH 4.5, respectively. Among the complex media tested, the malt extract medium and Phellinus igniarius medium were very good for mycelial growth of I. mikadoi. When Czapek-Dox medium was used as a minimal medium for cultivation of mycelia, xylose, raffinose and carboxymethyl cellulose were very excellent as a carbon and energy source. With respect to carbohydrate, sucrose and glucose were very good carbon sources. In general, organic complex nitrogen sources were better than other inorganic ones. As the organic complex nitrogen sources tested, yeast extract, soytone, proteose peptone and bacto peptone were the best as a source of organic nitrogen. When ammonium sulfate as an inorganic source of nitrogen was used, the enhanced mycelial growth was shown. p-Aminobenzoic acid was proved to be most appropriate source of vitamin. After the mycelia of I. mikadoi was cultivated at $27^{\circ}C$ for 5 days in MEM broth (pH 4.5), the activities of both exomycelial and endo-mycelial enzymes were determined. Among endomycelial enzymes assayed, the specific activity of laccase was much higher than those of other enzymes. When the fungus was grown in MEM broth, exomycelial specific enzyme activity of laccase was comparatively high. However, little or no enzyme activities of protease, chitinase and lipase were found.

• Journal of Embryo Transfer
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• v.10 no.2
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• pp.131-138
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• 1995

Essential and non-essential amino acids supplemented to culture medium stimulate mammalian embryo development in vitro. Amino acids such as glycine, taurine and alanine are concentrated in the lumen of oviduct and uterus and it can he thought that these amino acids may have physiological role on fertilization and embryo development. Our aim of this experiment was to investigate the effects of essential and non-essential amino acids, taurine or glycine supplemented to fertilization medium on the cleavage and subsequent in vitro development of bovine oocytes matured and fertilized in vitro. Immature oocytes were obtained from slaughtered Holstein cows and heifers and matured in TCM199 containing 10% fetal calf serum, 2.5 $\mu$g /mL of FSH and LH and 1 $\mu$g / mL of estradiol with granulosa cells in vitro. After maturation, oocytes were coincubated with sperm in fertilization medium supplemented with Minimum Essential Medium (MEM) essential and non-essential amino acids, taurine (3.75 mM) or glycine (10 mM) for 30 hours in vitro. Inseminated oocytes were cultured in synthetic oviduct fluid medium (SOEM) containing MEM essential, non-essential amino acids and 1 mM glutarnine up to 8 days after fertilization.Supplementation of fertilization medium with MEM essential and non-essential amino acids lowered significantly (p<0.05 and p<0.001) the cleavage rate after 30 hours of IVF (53.3%) and at Day 3 (62.7%: Day 0: the day of I VF) compared to control (64.3% and 77.3%, respectively). Subsequent developmental rates to morulae (Mo) and expanding blastocysts (ExBL) also significantly decreased (p<0.001 and p<0.05 for Mo and ExBL) when oocytes were coincubated with sperm in the medium containing MEM amino acids. Taurine added to fertilization medium have not increased the cleavage rate over the control, whereas glycine showed significantly lower (p<0.01) cleavage rate at Day 3 than that of taurine, but there was no significant difference in the developmental rates to Mo and ExBL of bovine embryos irrespective of the supplementation of taurine or glycine to fertilization medium. In conclusion, supplementation of fertilization medium with essential and non-essential amino acids, taurine or glycine has no beneficial effect on in vitro cleavage and development of bovine oocytes matured and fertilization in vitro.

• Journal of Periodontal and Implant Science
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• v.30 no.1
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• pp.39-52
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• 2000

Polypeptide growth factor belong to a class of potent biologic mediator which regulate cell differentiation, proliferation, migration and metabolism. 1,25-dihydroxyvitamin $D_3$ decrease cell proliferation, and stimulate alkaline phosphatase activity which express in osteoblast during cell differentiation period. IGF-I is known to stimulate cell proliferation and differentiation too. 1,25-dihydroxyvitamin $D_3$ is known to increase IGF-I binding sites and IGF binding protein which inhibite the effect of IGF. The purpose of this study is to evaluate potential role of IGF-I as mediator that control the action of 1,25-dihydroxyvitamin $D_3$. MC3T3-E1 cell were seeded $5{\times}10^5/ml$ at 100mm culture plate in ${\alpha}-MEM$ containing 10% fetal bovine serum. After 48 hour incubation period, medium were changed ${\alpha}-MEM$ containing 5% fetal bovine serum. After 24 hours, $10^{-9}M$ 1,25-dihydroxyvitamin $D_3$ added. Total mRNA was extracted at 0, 6, 24, 48, 72 hour. PRPCR method was programed for the detection of IGF-I mRNA. In the both groups of 1,25-dihydroxy vitamin $D_3$ treated and control, alternative splicing form of IGF-I, IGF-IA and IGF-IB were expressed. In the 1,25-dihydroxyvitamin $D_3$ treated group, IGF-I mRNA expression was matained until 24 hour, there after expression was decresed. MC3T3-E1 cell were seeded $2.5{\times}10^4/ml$ at 24well plate in ${\alpha}-MEM$ containing 10% fetal bovine serum. After 48 hour incubation period, medium were changed ${\alpha}-MEM$ containing 3% fetal bovine serum. After 24 hours, $10^{-9}M$ 1,25-dihydroxyvitamin $D_3$ and 10 ng/ml IGF-I were added separately or together. Cell were cultured for 1 and 3 days, $2{\mu}Ci/ml\;[^3H]$ -thymidine was added for the last 24h of culture of each days. ${[^3H]}$-thymidine incorporation in to DNA was measured and expressed counter per minute(CPM). DNA synthetic activity was significantly decreased by 1,25-dihydroxyvitamin $D_3$ both at 1 day and 3 day, and in the combination group of 1,25-dihydroxyvitamin $D_3$ and IGF-I, DNA synthetic activity was also decreased both at 1 day and 3 days. IGF-I did not affect the DNA synthetic activity compared to control group both at 1 day and 3 day. From the above results, 1,25-dihydroxyvitamin $D_3$ was potent inhibitor of cell proliferaton in MC3T3-E1 cells. It assumed that the effect of 1,25-dihydroxyvitamin $D_3$ on osteoblast proliferation may be mediated in part by decreased level of IGF-I.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2008.11a
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• pp.31-32
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• 2008

• Proceedings of the Optical Society of Korea Conference
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• 2004.07a
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• pp.258-259
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• 2004

• Journal of the Korean Society for Precision Engineering
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• v.25 no.12
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• pp.13-19
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• 2008