The biochemical composition of wild and cultured yellow croaker, Larimichthys polyactis, was analyzed in this study. The moisture contents in wild and cultured yellow croaker was high: $75.2{\pm}1.60%$ and $79.5{\pm}1.95%$, respectively. The crude lipid contents of wild and cultured yellow croaker were low; moreover, the crude protein and ash contents did nol differ significantly (P>0.05). The total amino acid content of wild and cultured yellow croaker did not differ significantly; however, the cystine content of wild yellow croaker was higher than than of cultured yellow croaker. The essential /nonessential amino acid (E/NE) ratio in wild and cultured yellow croaker was $0.76{\pm}0.01$ and $0.77{\pm}0.02$, respectively. The free amino acid and extractive nitrogen contents of cultured yellow croaker were high and differed significantly. The water soluble vitamin ($B_1$, $B_2$, $B_6$, $B_{12}$, C and folate) and fat-soluble vitamin (A and E) contents did not differ significantly. expect for niacin. The niacin content of cultured yellow croaker was higher than that of wild yellow croaker. The fatty acid composition of wild and cultured yellow croaker did not differ significantly The sodium, magnesium, and copper contents in wild yellow croaker were relatively low. In comparison, the calcium, phosphorus and iron contents in cultured yellow croaker were relatively high. Overall, the biochemical composition of wild and cultured yellow croaker did not differ significantly.
The effects of in vitro maturation and sperm treatment condition on the in vitro fertilization (IVF) and developmental capacity of bovine oocytes were investigated and the development of embryos was compared under the 2 different co-culture system, with GC or BOEC. The cultured embryo to 16 cell or morula wre transferred into recipients or frozen by 2 different freezing method. The results obtained were summarized as follows; 1. In vitro maturation rates of vovine follicular oocytes cultrued in TCM199 with 10% FCS or ECS were 64.0% and 72.7%, but the case of addition of 10% FCS or ECS to TCM199 co-cultured with granulosa cells were 81.3% and 84.0%, respectively. IVM rate of three TCM199 added to granulosa cells was higher than that of media without granulosa cells. 2. When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC and then fertilized in vitro by sperm treated with caffeine, embryo developments of bovine oocytes co-cultured with BOEC were 38.4% and 51.4%, respectively. But those of bovine oocytes co-cultured with GC were 52.2% by sperm treated with caffeine-heparin. 3. Cleavage rates of bovine oocytes cultured with 10% FCS alone and fertilized in vitro by sperm treated with caffeine-heparin was 33.0%. 4. When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC, embryo developments of bovine ooctyes co-cultured with BOEC of GC were 46.0% and 50.2%, respectively. 5. When bovine follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6. When Korean Native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developed co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6. When Korean Native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developments of the bovine oocyte co-cultured with BOEC and GC were 41.8% and 60.1%. But with FCS 10% those of the bovine oocytes co-cultured with BOEC and GC were 42.0% and 48.4%, respectively. 7. When Holstein's follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments fo the bovine oocytes co-cultured with BOEC and GC were 50.0% and 57.7%, but with ECS 10% those of the bovine oocytes co-cultured with BOEC and GC were 52.2% and 56.5%, respectively. 8. The viability of frozen-thawed embryos ranged from 60~80% and those of frozen-thawed embryos from vitrification was lower than that from conventional metiod. 9. The selected fresh embryos were transferred nonsurgically to 7 recipients but did not result in pregnancy.
This study was carried out to evaluate the cytotoxicity and anti-inflammatory effects of Resina Pini on cultured human gingival fibloblasts. We carried out a study of cytotoxic effects of Resina Pini on cultured cells by MTT assay. Various treatments on Resina Pini reduced its toxicity on cultured cells in order of natural Resina Pini, water extracted mixture of Resina Pini and Ramus Mori Albae and recrystalized Resina Pini. However, Resina Pini showed harmless levels of cytotoxicity to cultured human gingival fibroblast. (omitted)
Analyses of UV-Vis spectrum and reflectance are useful tools to identify pearls of which color was naturally made or chemically changed. Contents of some trace elements of pearls by using ED-XRF may give us the information that the pearls were originated from marine shells or freshwater shells, and may give us whether the pearls were chemically treated or not. Three types of chemically untreated pearls, freshwater cultured, Akoya cultured and Tahitian cultured, were treated with silver nitrate. UV-Vis absorbance spectrum of Ag was changed after treatment, and reflectance of pearls was decreased. Absorbance of natural black colored Tahitian cultured pearls at 400, 500 and 700 nm was observed. Natural black colored freshwater cultured pearls showed two characteristic absorption patterns in UV-Vis spectra at 380-400, 480-500 nm. Unlike naturally black colored Tahitian cultured pearls, the absorption spectrum at 700 nm could not be found from black freshwater cultured pearls. Manganese was not detected in the pearls from marine shells, and strontium content of the pearls from marine cultured shells was higher than that from freshwater cultured shells. According to ED-XRF analysis, Ag contents of silver nitrate treated pearls increased as more time passed, because silver ion invaded to nacre layer of the pearls. Since Ag content of the black pearls known as untreated ranged under 0.1%, analysis of Ag content may used to distinguish the black pearls are treated or not in marketplace.
Mammalian oviductal epithelial cells have been known to improve in vitro fertilization and embryonic development. Recently, co-cultured human embryos with the epithelial cells in human genital tract has been reported to improve the pregnancy rate. The purpose of the study was to investigate the effects of the epithelial cells of human genital tract on the development of mouse early embryos and human fertilized oocytes. The epithelial cells of human genital tract were collected from the fallopian tubes which were obtained during hysterectomy in fertile women and from the endometrium during endometrium biopsy. Collected human ampullary cells(HACs) and endometrial cells(HECs) were cultured for 10 days to establish primary monolayer. Second passaged HACs and HECs were obtained by trypsinization were cryopreserved in PBS with 1.5 M DMSO for later use. To investigate the effect when co-cultured with HACs and HECs, we tried to apply strict quality control on mouse embryo, from two cell to blastocyst prior to human trial. The results of quality control were as follows; In Group I (Ham's F10 with 10% FCS), Group IT (co-cultured with HACs) and Group ill (co-cultured with HECs), developmental rates to blastocyst were 63.3%(253/400), 76.0%(304/ 400),74.0%(296/400), respectively. Hatching rates were 36.8%(147/400), 41.80/0(167/400), 38.0%(152/400), respectively(p<0.05). To perform the human IVF, cryopreserved HACs were thawed at 37$^{\circ}C$ waterbath, seeded on the well dish and cultured for 48 hI'S. The pronuclear stage embryos were transferred to the seeded well dish. After 24 hRS, co-cultured embryos were examined and transferred to patient's uterus. The results of human IVF when co-cultured with HACs were that fertilization and developmental rates were 61.8% (256/414), 95.3% (244/256) as compared with 57.2% (279/488) and 94.6%(264/279) in Ham's F10 supplemented with 10% FCS(control). However, 62.9% (161/256) of co-cultured human embryos showed good embryos(no or slight fragmentation) as compared with 53.8 % (150/279) in control(p < 0.05). Pregnancy rate was 40.0% (12/30) when co-cultured with HACs whereas 30.6%(11/36) in control. In conclusions, co-culture system using HACs and HECs improved the developmental and hatching rates of mouse embryo. Also, in human IVF system when co-cultured with HACs, it improved both the quality of human embryos and the pregnancy rate.
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.
These studies were carried out to investigate the effects of fetal calf serum(FCS), estrous porcine serum(EPS), porcine follicular fluid(PFF), hormone and matured cumulus cell(MCC) on in vitro maturation and fertilization of porcine follicular oocytes. The ovaries and testes were obtianed from slaughtered Landrace sow and boars, respectively. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluid from the visible follicles of diameter 3~5 mm and the semen were prepared from boar's epididymal cauda. The follicular oocytes were cultured in TCM-199 medium containing hormones, FCS, EPS, PFF and MCC for 48hrs. in a incubator with 5% CO2 in air at 36$^{\circ}C$ and then matured oocytes were again cultured for 18~20 hrs. with $1.5\times$106/ml motile capacitated sperm in the modified Tyroide solution containing 100$\mu\textrm{g}$/ml of heparin. The results obtained in these experiments were summarized as follows : 1. The maturation and fertilization rate of the follicular oocytes, cultured in the TCM-199 medium supplemented with 10% FCS and PMSG+HCG were 55.6~64.5% and 33.3~37.1%, respectively. 2. The maturation and fertilization rate of the follicular oocytes cultured in the TCM-199 medium supplemented with 20% EPS and PMSG+HCG were 50.0~55.0% and 30.3~33.3%, respectively. 3. The maturation rate(59.0~64.2%) and fertilization rate(34.8~39.3%) of follicular oocytes cultured in TCM-199 medium supplemented 20% FCS and 50% PFF were higher than those of follicular oocytes cultured in TCM-199 medium supplemented with 5%, 10% and 15% FCS and 10% and 50% PFF. 4. The maturation rate(60.0%) and fertilization rate(40.0%) of follicular oocytes cultured in TCM-199 medium supplemented with 20% FCS and granulosa cell (1$\times$106/ml) were significantly higher than those of fiollicular oocytes cultured in TCM-199 medium supplemented with 5%, 10% and 15% FCS and granulosa cell.
This study was carried out to investigate on the survival rates and in vitro developmental rates of bisected bovine embryos were by manipulator and micropipette. Bisected embryos were co-cultured in 20% FCS(v/v)+TCM-199 media containing hormones, oviductal epithelial cells and cumulus cells 0 to 72 hrs after bisection. Survival rate and in vitro fertilization rate were defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The in vitro developmental rate of biseciton embryos co-cultured in 20% FCS+TCM-199 medium containing PMSG, hCG, PMSG+hCG, hCG+$\beta$-estradiol 0 to 20 hrs and 20 to 40 hrs were 36.7, 26.7, 33.3, 40.0, and 30.0, and 30.0, 33.3, 30.0, 26.7, and 26.7%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing hormones was significantly higher than that of non co-culture(25.0%). 2. The in vitro developmental rates of bisection embryos co-cultured in 20% FCS+TCM-199 medium containing oviductal epithelial cells 4 to 5 hrs and 20 to 24 hrs were 40.0 and 33.3%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing oviductal epithelial cells was significantly higher than that of non co-culture(25.0%). 3. The in vitro developmental rates of bisection embryos co-cultured in 20% FCS+TCM-199 medium containing cumulus cells 4 to 5 hrs and 20 to 24 hrs were 43.3 and 36.7%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing cumulus cells was significantly higher than that of non co-culture (25.0%).
This study investigated the seasonal changes of body composition and elasticity between wild and cultured brown croaker, Miichthys miiuy. The wild fish were analyzed and compared with cultured fish in moisture, crude protein, lipid, ash and many kinds of amino acids. Cultured fish was higher in moisture content and lower in crude lipid and protein content than those of wild one. The wild fish were more abundant in the total amino acid compositions than those of cultured one. As result E/A ratio there was a little significant differences between wild and cultured. In highly unsaturated fatty acid, EPA (Eicosapentaenoic Acid) and DHA (Docosahexaenoic Acid) content of cultured fish were higher than wild one. On the other hand, the gel strength, max weight and hardness of wild fish were higher than cultured one.
This study examined Tetraselmis sp. JK-46 isolated from seawater from the East Sea. Deep seawater (DSW) had a greater effect on the growth of Tetraselmis sp. JK-46 than surface seawater (SSW). The crude protein, lipid, carbohydrate and ash contents of Tetraselmis sp. JK-46 cultured with DSW were 27.2, 37.1, 13.2 and 26.3 %, respectively, and these values were similar to the results for samples cultured with SSW. The contents of Mg, Ca, Fe and K in the DSW cultured samples were 7080.3, 1009.6, 251.2, and 2749.7 mg/100 g, respectively. The fatty acid compositions of Tetraselmis sp. JK-46 cultured with DSW and SSW were 53.7 and 49.0 % polyunsaturated fatty acids (PUFA) and 25.7 and 30.7 % saturated fatty acids (SFA), respectively. The total amino acid contents of the samples cultured with DSW and SSW were 7392.6 and 6376.0 mg/100 g respectively. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of Tetraselmis sp. JK-46 extracts increased with the concentration of the chloroform and ethyl acetate fractions. The half maximal inhibitiory concentrations ($IC_{50}$) of the chloroform and ethyl acetate fractions of DSW and SSW cultured samples were 1.2 and 2.6 mg/mL, and 3.1 and 3.3 mg/mL, respectively. The ethyl acetate fractions of DSW and SSW cultured samples has anticoagulant activity and the activated partial thromboplastin times (APTT) were 93.4 and 89.3 sec., respectively. The chloroform and ethyl acetate fractions showed antimicrobial activity against Bacillus subtilis, Escherichia coli and Candida albicans.
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