• Biomolecules & Therapeutics
• /
• v.3 no.2
• /
• pp.116-121
• /
• 1995

Some multidrug resistance inhibitors have been known to be influenced by the serum concentration. In this study, effect of serum concentration on inhibition of nucleoside transport by BIBW22, a new multidrug resistance inhibitor derived from dipyridamole (DPM), was studied. When 5% or 100% (v/v) of fetal bovine serum (FBS) was contained in the culture, DPM dose for which nucleoside transport was inhibited by 50% (lD$_{50}$) was 0.42 $\mu$M or 1.17 $\mu$M, respectively. BIBW22 also showed the same trend as DPM did in response to increase of FBS concentration. However, ID$_{50}$ value for DPM in the absence or presence of human plasma was 0.007 $\mu$M or 1.02 $\mu$M respectively showing 145 times increase of ID$_{50}$ value. ID$_{50}$ value for BIBW22 in the presence of human plasma was 0.028 $\mu$M showing only 5 times increase in ID$_{50}$ value. This result suggests that potency of BIBW22 was much less affected by the plasma concentration and BIBW22 could be a good candidate for a clinical use in multidrug resistance treatment.treatment.

• Journal of Veterinary Clinics
• /
• v.9 no.1
• /
• pp.323-332
• /
• 1992

The present study was carried out to examine the effect of oviduct epithelium and its conditioned medium on e development of early bovine embryos in vitro. Oocytes obtained from ovarian follicles of slaughtered cows were cultured in TCM199 with 10% fetal calf serum for 22-24hrs and then fertillzed in vitro using frozen-thawed semen treated with BO-caffein, BO-BSA(20mM heparin added). Oviduct epithelium was collected in each stage of the estrus cycle and conditioned medium was the medium in which oviduct epithelium in early luteal stage was cultured. In vitro fertilized bovine embryos of 1～2 cell were co-cultured with oviduct epithelium from different estrus cycles, cultured in conditioned medium, and cultured in rabbit oviduct. The cleavage rates of in vitro fertilized early bovine embryos co-cultured with oviduct epithelial cell from early luteal, luteal and follicular phase of estrus cycle(67.2～70.8%) and cultured in conditioned medium(56.7%) were significantly(p<0.05) higher than that of the control(44.2%) The rate of development to morula or blastocyst stage in oviduct epithelial cell co-culture(15.3～32.5%) from three phase of estrus cycles and conditioned medium(14.5%) were significantly(p<0.05) higher than that of the control(5.2%). The oviduct epithelial cell from early luteal phase gave a significantly( p<0.05) higher rate of development to morula or blastocyst stage than both luteal and follicular phase. The results of in vivo culture in rabbit oviduct of early bovine embryos were 52.1% for the cleavage rate and 26.7% for the rate of development to morula or blastocyst stage.

• Korean Journal of Animal Reproduction
• /
• v.18 no.2
• /
• pp.87-94
• /
• 1994

The objective of this study is to evaluate the developmental ability of mouse embryo in the presence of human amniotic fluid (hAF), The highest development rate was found in the culture media supplemented with 20% mid-term hAF but this rate was concomitantly reduced with more than 20% hAF. Furthermore, mouse two-cell embryos cultured in 20% mid-term hAF were developed more consistently to the expanded and hatched blastocyst stages compared to those cultured in simple medium. However, no significant differences in the embryo development rates were observed among the supplemented effects of 20% mid-term hAF, 0.3% bovine serum albumin (BSA), and 10% fetal calf serum (FCS), Development rates of two-ceiI mouse embryos cultured in 20% full-term hAF were declined compared to 20% mid-term hAF. Outgrowth of hatched blastocysts were observed when the embryos were cultured in medium containing 20% mid-term hAF or 10% FCS. But two-cell mouse embryos cultured in the presence of 20% full-term hAF or O.3% BSA was not observed their outgrowth. The kinetics of outgrowth processes in the presence of hAF were similar to those with 10% FCS. However, embryos with FCS showed a considerably greater extents of trophetodermal cell proliferation and outgrowth. Taken together, these data suggest that mid-term hAF may have a suitability for the mammalian embryos and induce embryonic outgrowth.

• The Korean Journal of Zoology
• /
• v.34 no.2
• /
• pp.148-158
• /
• 1991

The purpose of this research was to establish the culture condition for dissociated submandibu -lar gland (SG) cells. After trypsin digestion of SG from 3-4 weeks old mice, dissociated cells were cultured in 1OO/o fetal bovine serum-Dulbecco's modified Eagle's medium (FBS-DME) or 0.5-2% low protein serum replacement-DME (LPSR-DME) on plastic surface to form monolayer. The effects of FBS, LPSR and hormones on the growth and function of cultured SG cells were examined. SG cells dissociated by enzyme were successfully cultured and were characterized as epithelial-like cells by light and electron microscope. The maximal DNA synthesis of cultured SG cells was achieved by DME containing 5-10% FBS. The same results were obtained when the effects of LPSR on cell proliferation were examined up to a LPSR concentration of 2%. SG cells cultured in 20/o LPSR-DME expressed a population doubling time of 42.5 hrs and a saturation density of 1.2 $\times$10 5cell/cm$^2$. Dihydrotestosterone (DHT) in medium did not influence on the DNA synthesis of the cultured SG cells, but stimulated protein synthesis of the SG cells. Thyroxine (T4) stimulated protein synthesis of the SCI cells markedly in a dose-dependent fashion. EGF secretion by the cultured SG cells increased significandy by DHT and or T4 trearment. This finding indicated that secretion of EGF by the SG cells was under the control of the hormones such as androgen and thyroid hormones. It seems to be that the culture condition described here can be used as a useful tool for further research on the SCI cells.

• Korean Journal of Animal Reproduction
• /
• v.16 no.1
• /
• pp.55-62
• /
• 1992

Bovine oocytes obtained from ovarian(2 to 5mm in diameter) of slaughtered cows were cultured in TCM 199 with 10~20% estrous-cow-serum(ECS) for 24~25hr at 39$^{\circ}C$ in 5% CO2-95% air. After culture, some oocytes were examined their maturation. The remainder were used to assess the fertilizability with frozen-thawed spermatozoa in a medium containing caffeine and heparin, and subsequent development in media with bovien cumulus cells(BCC) or bovine oviduct epithelial cells(BOEC). The results obtained were summarized as follows ; 1. The maturation rate of the oocytes in TCM199 with 15% ECS group(76.5%) was higher than that of 10% ECS(69.2%) or 20% ECS group(64.8%). 2. The proportions of the oocytes penetrated and the pronuclear oocytes in the presence of caffeine and heparin were 72.1%(62/86) and 93.5%(58/62), respectively. The rate of polyspermy in the fertilized oocytes was 8.1%. 3. When 73 oocytes recovered from fertilization drop were cultured in TC-199 medium with 10% fetal calf serum(FCS), 41 oocytes(56%) cleaved to 2-cell and further stages of embryos. Among these only one embryo developed upto morula stage. 4. The rate of the cleaved oocytes was higher in medium with BCC(80%:59/74) than BOEC(76%:58/76). However, the rate of developed morulae and blastocysts was higher in the medium with BOEC(40%;23/58) than with BCC(34;20/59).

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.5 no.1
• /
• pp.44-47
• /
• 2000

Cell growth and DNA synthesis were studied from a cultured early- and late- pas- sage mouse aorta smooth muscle cell (MASMC) because the proliferation of vascular smooth muscle cell (VSMC) is a key factor in development of atherosclerosis. In this study, the cells were cultured in fetal bovine serum (FBS) and stimulated by growth factors such as thrombin and platelet-derived growth factor-BB (PDGF-BB). Compared to the number of early-passage MASMC (passage 3 to 9) the number of late-passage MASMC (passage 30 to 40) in a normal serum state was increased 2 fold at Day 1, 3 and 6 in culture, respectively. Incorporation of $[^3H]$ thymidine into DNA induced by serum, PDGF and thrombin in late-passage MASMC was greater than those in early-passage MASMC. We also examined whether intracellular zinc levels would be an aging factor or not. The intracellular zinc level in early- and late-passage MASMC was monitored by using the zinc probe dye N-(6-methoxy-8-quinolyl)-p-toluenesulfonamide. It is interested that late-passage MASMC increased the intracellular fluorescence level of zinc, more than the early passage MASMC did. The alterations of intracellular zinc level occur concurrently with changes in MASMC proliferation rate during aging. This data suggest that the age-associated changes in zinc concentrations may provide a new in vitro model for the study of smooth muscle cell differentiation.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.26 no.6
• /
• pp.1194-1199
• /
• 1997

The effect of hot taste preference on selected immune responses was investigated in human peripheral immunocompetent cells. Human lymphocytes and natural killer(NK) cells were prepared at a concentration of 2$\times$$10^{6}$ cells/ml in RPMI-1640 containing 10% fetal bovine serum. Lymphocytes proliferation was determined with the [$^{3}H$]-thymidine pulse for 18hrs after concanavalin A, phytohemagglutinin, Salmonella typhimurium mitogen, or media alone. NK cell activity was measured by cytolysis of $^51Cr$-labeled target cells K562. Serum antibodies levels such as IgM, IgG, IgA were also measured by ELISA method. There was no difference of serum IgM level among the groups, but IgG and IgA levels were greater in the group with hot taste preference than those of the group without hot taste preference. In lymphocytes of the group with hot taste preference there was a greater mitogen-induced lymphocyte proliferative responses compared to the group without hot taste preference. In addition, NK cell activity in group with hot taste preference was lower than that of the group without hot taste preference. These results suggest that the eating habit of spicy food containing hot components may affect immune status by modulating selective immunocompetent cells function.

• BMB Reports
• /
• v.54 no.7
• /
• pp.374-379
• /
• 2021

Tetranectin (TN), an adipogenic serum protein, enhances adipocyte differentiation, however, its functional mechanism has yet to be elucidated. In the present study, we investigated the adipogenic function of TN by using medium containing TN-depleted fetal bovine serum (TN-del-FBS) and recombinant mouse TN (mTN). The adipocyte differentiation of 3T3-L1 cells was significantly enhanced by mTN supplementation essentially at differentiation induction, which indicated a potential role of the protein in the early differentiation phase. The adipogenic effect of mTN was more significant with insulin in the differentiation induction cocktail, implicating their close functional relationship. mTN enhanced not only the proliferation of growing cells, but also mitotic clonal expansion (MCE) that is a prerequisite for adipocyte differentiation in the early phase. Consistently, mTN increased the phosphorylation of ERK in the early phase of adipocyte differentiation. Results of this study demonstrate that the adipogenic function of mTN is mediated by enhancing MCE via ERK signaling.

• Korean Journal of Animal Reproduction
• /
• v.17 no.4
• /
• pp.271-278
• /
• 1994

The aim of this study was to compare the two culture systems 1) co-culture with cumulus cells and 2) chemically defined medium supplemented with amino acids (CR1aa) and fetal calf serum (FCS) of in vitro produced bovine embryos from follicular oocytes in vitro. Bovine follicular oocytes were collected from ovaries of slaughtered cows and matured in TCM199 supplemented with 10% FCS and hormones (1$\mu\textrm{g}$/ml FSH-P and 1$\mu\textrm{g}$/ml oestradiol-17$\beta$)24 hours at 39$^{\circ}C$ under 5% CO2 in air. The capacitation of spermatozoa from ejaculated or frozen bull semen was induced by centrifugation through Percoll density gradient (45%, 90%). Then capacitated spermatozoa (1$\times$106/ml) were inseminated into 50${mu}ell$ droplet containing matured follicular oocytes and incubated for 40~42 hours. Cleaved embryos of 2~4cell stage were transferred to the co-culture with cumulus cells and/or CR1aa medium supplemented with FCS. In semen source, the developmental rates to the blastocyst and the hatched blastocyst stages were higher in ejaculated semen(27.6% and 14.9%) than those of frozen-thawed semen(18.3% and 11.8%), respectively. In two culture systems, the proportions of embryonic development upto the blastocysts and the hatched blastocysts were higher of CR1aa medium (22.1% and 12.1%) than those of cumulus cell co-culture (16.8% and 5.1%), respectively. The number of cells in exapnded blastocysts was slightly higher in cumulus cells co-culture (122.6$\pm$8.5) than that in CR1aa medium (117.9$\pm$5.9). The present results indicated that the early development of in vitro produced bovine embryos can be maintained efficiently in CR1aa medium as well as in co-culture with cumulus cells.

• Korean Journal of Animal Reproduction
• /
• v.24 no.1
• /
• pp.49-57
• /
• 2000

This study was conducted to investigate the developmental potential of cloned embryos derived from bovine fetal fibroblast cells, and the effect of quiescent treatment, passage number and origin of donor cells on in vitro development of cloned embryos. Fetal skin and liver-derived fibroblast cells were transferred to enucleated oocytes after serum starvation or nontreatment (cycling). After electrofusion. reconstituted embryos were activated with $Ca^{＋＋}$-ionophore and cycloheximide, and cocultured for 7~9 days with BRL cells. Some blastocysts were transferred to recipient cows 7~8 days post estrus. The development rate to the blastocyst stage of serum starved cell-derived embryos was higher (25.3%) than that of actively dividing cells-derived embryos (15.9%), The rates of blastocyst formation were 23.1~25.0% after transfer of cell passaged 4 to 6 times, and 23.8 and 25.2% after transfer of fetal skin and liver cells, respectively. After embryo transfer, 34.4% and 15.6% of recipient cows were pregnant on Day 60 and 120, respectively, and one male calf was produced from skin-derived vitrified blastocyst. The result of this study showed that the development of cloned embryos. was enhanced by quiescent treatment, but did not different among the cells passaged 4 to 6 times, and between skin and liver cells. This result also confirms that offspring can be obtained from the vitrified clone embryo derived from fetal skin cell.