• Title/Summary/Keyword: Feeder layer

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Systems for Production of Calves from Cultured Bovine Embryonic Cells (우 수정란의 배양세포들로부터 송아지 생산을 위한 체계)

  • ;N. L. First
    • Korean Journal of Animal Reproduction
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    • v.18 no.4
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    • pp.299-307
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    • 1995
  • The goal of cell stem cell technology is to produce a viable and genetically normal animal. To achieve this goal various laboratories have followed 2 different pathways beginning with either the culture of 1) single or pooled ICMs grown with or without a feeder layer or 2) single or pooled 16-20 cell stage embryos grown with a feeder layer. Also, thus far embryonic cell cultures or lines have been established by several methods including loose suspension culture for short-term cultures and more commonly murine or bovine fibroblast feeder layers for long-term culture. Pluripotent lines have been derived from 16-cell through blastocyst inner cell mass stages. The efficiency of establishing cell lines and cell proliferation apper to be affected by the number of cells or embryos starting the line. Most attempts to produce offspring from long term STO cell feeder layer cultured ICM or morulae derived ES cells have resulted in pregnancy failure in the first trimester when ES cells were used in cuclear transfer or have failed to retain ES cells in the progeny produced by chimerization. The exception is 1 chimeric fetus from use of morula ES cells in the chimerization with early embryonic cells. There is much to be learned yet about ES cell culture requirements for maintenance of totipotency. If bovine ES cell lines loose imprinting pattern and totipotency with long-term culture and passage as suggested for mouse ES cells, we may be limited to the use of short-term cultures for multiplication of embryos and efficient production of transgenic animals. No bovine ES cell system has yet met all of the criteria indicated for a totipotent ES cell line.

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Density Composition and Feeding Guild of the Dominant Polychaetous Community in Shallow Muddy Bottom in Tomioka Bay, Amakusa, Japan

  • LEE Si-Wan;PAIK Eui-In
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.6
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    • pp.793-804
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    • 1995
  • Polychaetous community survey in Tomioka Bay was carried out 5 times seasonally from May 1991 to March 1992 by quantitative grab sampling (0.05m2) at 11 stations. Based on the granulometric composition and environmental factors, a homogeneous soft bottom was found in St.5-10. The species of the polychaete were classified into three feeding groups using the Fauchald and Jumars' feeding guild system. According to polychaetous community composition data, deposit feeders predominate in sandy silt area where the silt-clay content is $60-69.3\%.$ These deposit feeders were subdivided into surface deposit feeders and subsurface deposit feeders by their living position and mode. Also, suspension feeding group comes as the third dominant group. Seasonal changes of each feeding group were described in terms of numerical density and biomass. Feeding layer and types of dominant species (Lumbrineris longifolia: surface deposit feeder; Praxillella pacifica: subsurface deposit feeder; Chone duneri; suspension filter feeder, etc.) were examined in the intact sediment core samples. Also, longterm density change among the three dominant species during 10 years was disussed.

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Efficient isolation, culture and regeneration of Lotus corniculatus protoplasts

  • Raikar, S.V.;Braun, R.H.;Bryant, C.;Conner, A.J.;Christey, M.C.
    • Plant Biotechnology Reports
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    • v.2 no.3
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    • pp.171-177
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    • 2008
  • This paper reports an improved protocol for isolation, culture and regeneration of Lotus corniculatus protoplasts. A range of parameters which influence the isolation of L. corniculatus protoplasts were investigated, i.e., enzyme combination, tissue type, incubation period and osmolarity level. Of three enzyme combinations tested, the highest yield of viable protoplasts was achieved with the combination of 2% Cellulase Onozuka RS, 1% Macerozyme R-10, 0.5% Driselase and 0.2% Pectolyase. The use of etiolated cotyledon tissue as a source for protoplast isolation proved vital in obtaining substantially higher protoplast yields than previously reported. Culture of the protoplasts on a nitrocellulose membrane with a Lolium perenne feeder-layer on the sequential series of PEL medium was highly successful in the formation of microcolonies with plating efficiencies 3-10 times greater than previous studies. Shoot regeneration and intact plants were achieved from 46% of protoplast-derived cell colonies.

A study on design for the $\pi$-junction of a feeder waveguide with an inductive wall using FDTD method (FDTD법을 이용한 유도성벽을 가지는 $\pi$분기 급전도파관의 설계에 관한 연구)

  • 민경식;김광욱;고지원;김동철;임학규
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2000.05a
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    • pp.143-146
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    • 2000
  • This paper presents a study on design for the k-junction of a feeder waveguide with an inductive wall using FDTD method. The feed structure consists of a single waveguide plated on the same layer as radiating waveguide and is characterized by the unit divider, railed a $\pi$-Junction. This $\pi$-Junction with an inductive wall splits part of the power into two branch waveguide through one coupling window, and can excite densely arrayed waveguide at equal phase and amplitude. The power dividing characteristics of a $\pi$ -Junction obtained by FDTD method are compared with one of Galerkin's method of moments. The scattering matrices a $\pi$ -junction calculated by FDTD method are realized.

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Maintenance and Differentiation of Pluripotential Embryonic Cell Lines from Mouse Blastocysts (BCF1 생쥐 배반포기 유래 배아간세포 작성에 관한 연구)

  • 이재원;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.18 no.4
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    • pp.235-244
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    • 1995
  • The present study was designed to demonstrate that ES cell lines efficiently could be isolated from explanted blastocysts of hybrid BCF1 mouse when grown on STO feeder layer derived from mouse fibroblasts in culture medium supplemented with leukemia inhibitory factor (LIF). The expanded blastocysts were attached to mitomycin C-inactivated STO feeder layer and were cultured for 4 days. Four days later the ICM was disaggregated by a short term trypsin treatment (0.25% trypsin / 0.04% EDT A for 2-3 min). The resulting cell suspension was seeded on a new STO feeder layer and covered with DMEM supplemented with 10% FCS, 0.1 mM nonessential amino acid, 0.1 mM sodium pyruvate, 0.1 mM mercaptoethanol and 1,000 U/ml LIF. Colonies of ES-like cells were observed after the second passage. These colonies were repeatedly passaged at approximately 5 day intervals. In this study, five ES-like celllines were isolated by directly explanting blastocysts, but three lines were lost after the 5th passage, possibly due to toxic effects of a new FCS batch. The characterization of developmental potential of isolated cell lines was performed with respect to in vitro differentiation and specific activity of alkaline phosphatase (AP). When cells were cultured in suspension, the aggregates of cell lines were capable of forming simple embryoid bodies (EB), and showed the capacity for forming cystic multilayer EBs. In addition, the cell lines were positive for AP staining, a biochemical marker characteristic of mouse ES cells.

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Effect of the Isolation Method of Mouse Inner Cell Mass, Types of Feeder Cells and Treatment Time of Mitomycin C on the Formation Rate of ICM Colony (생쥐 내세포괴의 분리방법과 지지세포의 종류와 Mitomycin C 처리 시간이 내세포괴 Colony 형성률에 미치는 영향)

  • Jang, Ho-Jin;Ko, Kyung-Rae;Kim, Mi-Kyung;Na, Yong-Jin;Lee, Kyu-Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.4
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    • pp.265-272
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    • 2006
  • Objective: This study was carried out to evaluate the effect of the isolation methods of inner cell mass from mouse blastocyst, types of feeder cells and treatment time of mitomycin C on the formation rate of ICM colony. Methods: The inner cells were isolated by conventional immunosurgery, partial trophoblast dissection with syringe needles and whole blastocyst co-culture method. Commercially available STO and primary cultured mouse embryonic fibroblast (pMEF) feeder cells were used, and mitomycin C was treated for 1, 2 or 3 hours, respectively. The formation rate of ICM colony was observed after isolation of ICM and culture of ICM on the feeder cells for 7 days. Result: The ICM colony formation rate on STO were significantly higher in partial trophoblast dissection group (58%) than that in immunosurgery (12%) or whole blastocyst culture (16%) group (p<0.05). The formation rate on pMEF feeder layer was higher in partial trophoblast dissection (88%) and whole blastocyst culture (82%) group than that in immunosurgery (16%) group (p<0.05). When mitomycin C treated to pMEF for 2 hours, the formation rate of 88% was significantly higher than those of other conditions. Conclusion: Above results showed that the efficient isolation method of ICM from blastocyst was the partial trophoblast dissection and the appropriate treatment time of mitomycin C was 2 hours. However, the subculture of ICM colony and characterization of stem cells should be carried out to confirm the efficacy of the partial trophoblast dissection method.

Design of Plano-Convex Lens Antenna Fed by Microstrip Patch Considering Integration with Microwave Planar Circuits

  • Yu, Seung-Gab;Yeon, Dong-Min;Kim, Yong-Hoon
    • Journal of electromagnetic engineering and science
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    • v.1 no.1
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    • pp.67-72
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    • 2001
  • In this paper, the plano-convex lens antenna fed by a single patch is studied for a microwave remote-traffic monitoring sensor with constraints of small size and low cost. Measurement of an AUT (Antenna Under Test) involves the considerations of a triangular groove for matched layer and metallic shielding effects. A formulation for extracting the parameters of a piano-convex lens antenna, based on geometrical optics, is introduced using Fermat`s principle of the equi-phased ray condition. Teflon ($\varepsilon_{{\gamma}}$/ =2.0) is chosen as a material of a plano-convex lens antenna for adjustment of aberrations on the lens surfaces automatically. A fabricated plano-convex lens shows 3-dB beamwidth of 7.5 degree and side-lobe level of -29 dB with an aperture distribution of the parabolic-squared taper on pedestal. This lens supports easier integration with the planar microwave circuits by using a microstrip single patch as a primary feeder of the lens antenna.feeder of the lens antenna.

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Protein Profile of Embryonic Stem Cells Following Differentiation

  • D. I. Jin;Kang, H. S;Kim, H. J.;Lee, S. H.;Park, C. S.;K. S. Im;Lee, H. M.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.377-384
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    • 2002
  • To investigate the expression patterns of proteins and growth factor signals in differentiated rabbit embryonic stem (ES) cells, ES cells with confluent stage grown of feeder layer and differentiated cells into embryoid bodies (EB) without feeder cell were applied to protein gel and Western blotting analysis. There were 66kDa and 28kDa specifically expressed in differentiated ES cell but not in undifferentiated ES cell while 25kDa protein band showed up in only undifferentiated ES cells. Also there were some difference of protein bands in several area of gel between differentiated and undifferentiated ES cells such as about 100 kDa, 50kDa and 27kDa areas, but there was no difference in band pattern of one-dimensional gel analysis between mouse ES cells and rabbit ES cells. IGF-I receptor and EGF receptor were expressed in differentiated cells and undifferentiated cells. And ICF-I and EGF were not expressed in both differentiated and undifferentiated cells. These results indicated that ES cells express their own proteins to inhibit differentiation while EB cells synthesize different proteins to differentiate, and 16F-I receptor and EGF receptor were expressed in both ES and EB cells probably for the different functions.