• Title/Summary/Keyword: Animal cell

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The Influence of Microinjection of Foreign Gene into the Pronucleus of Fertilized Egg on the Preimplantation Development, Cell Number and Diameter of Rabbit Embryos

  • Makarevich, A.V.;Chrenek, P.;Fl’ak, P.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권2호
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    • pp.171-175
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    • 2006
  • The aim of this in vitro study was to test the effect of microinjection (Mi) of foreign gene into the rabbit egg pronucleus and epidermal growth factor (EGF) addition on the blastocyst rate, the cell number and the diameter of embryos, and to determine possible relationships between embryo cell number and embryo diameter. Blastocyst rate was significantly decreased in gene- Mi (G-Mi/E0) group (63.1%) comparing to intact ones (83.5%, $p_1$<0.05). The addition of EGF at 20ng/ml (G-Mi/E20) or 200 ng/ml (GMi/ E200) to gene-Mi embryos did not affect blastocyst rate (65.6 and 55.2% resp.). As a control for Mi, the eggs were microinjected with the same volume of phosphate-buffered solution (PBS-Mi) instead of the gene construct solution. Cell numbers and embryo diameters were measured from embryo images obtained on confocal laser scanning microscope. Bonferroni-modified LSD test showed that the embryo cell number in PBS-Mi group was significantly lower ($p_1$<0.05) and in gene-Mi group was tended to decrease compared with intact embryos. Embryo diameter was not different among experimental groups. No effect of EGF given at any doses both on the cell number and embryo diameter was found. A positive correlation between cell number and embryo diameter was observed in all groups of embryos. Since embryo diameter was not changed under the influence of Mi or EGF addition in this study, this seems to be more conservative characteristics of the embryo morphology. These results suggest that the pronuclear microinjection compromises developmental potential of embryos, decreasing blastocyst rate and embryo cell number, whilst embryo diameter is not affected. No effects of EGF on studied parameters were confirmed. Declined quality of Mi-derived embryos is caused by the microinjection procedure itself, rather than by the gene construct used.

C6 Rat Glioma Cell에서 리튬에 의한 성장 억제와 Insulin-like Growth Factor System Components의 발현과의 관계 (Relationships of the Lithium-Induced Growth Inhibition of C6 Rat Glioma Cell to Expression of the Insulin-like Growth Factor System Components)

  • 김인애;진은정;조은정;손시환;이철영
    • Journal of Animal Science and Technology
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    • 제46권4호
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    • pp.563-570
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    • 2004
  • Insulin-like growth factors(IGF)s-I과 -II ligands와 이들의 receptors 및 6종류의 IGF-binding proteins(IGFBPs)로 구성된 IGF systim은 여러 종류의 세포의 생존, 증식 및 분화에 있어서 매우 중요한 역할을 한다. 리튬은 in vitro에서 여러 종류의 세포의 생존과 증식의 조절제로 알려져 있다. 본 연구는 IGF-I, IGF-I receptor 및 IGF carrier로서 주로 IGFBP-3를 발현하는 rat C6 glioma cell에서 LiCl로 유도된 세포 생존 및 증식의 변화와 IGF system components의 발현간의 관계를 구명하고자 수행되었다. 10%혈청을 함유하는 배양액에서 0, 2mM, 혹은 5mM LiCl을 첨가하여 C6 cell을 24시간 배양했을 시 세포의 생존률과 세포 수는 리튬 첨가에 의해 영향을 받지 않았다. 그러나 72시간 배양 했을 때 C6 cell은 명백히 리튬의 첨가수준에 따라 반응하였다. 0, 2m, 5mM LiCl 첨가수준에서 72시간 배양한 C6 cell은 각각 전형적인 세포분열, 세포분열 중지 및 세포사멸 양상을 보였다. 더욱이 사멸돼가는 세포는 reverse transcription-polymerase chain reaction으로 조사한 IGF-I, IGF receptor 및 IGFBP-3의 발현수준이 저하되었다. 흥미롭게도 혈청을 첨가하지 않은 배양조건 하에서 IGFBP-3에 대한 antisense oligodeoxyribonucleotide를 10${\mu}M$ 수준의로 첨가하여 배양했을 때도 표적 mRNA는 물론 세포 수도 줄었다. 종합하자면, C6 cell에서 리튬의 독성 효과의 일부는 이 제제에 의한 IGF system components의 발현 억제 효과에 의해 매개될 소지가 크다. 이러한 관점에서 IGRBP-3는 적어도 이 세포의 정상적인 증식을 위해 꼭 필요한(‘prrmissive') 역할을 할 수 있다는 점을 시사한다.

산양유의 특성 - 유지방, 체세포, 그리고 산양취 - (Characteristics of Goat Milk - Milk Fat, Somatic Cell Count, and Goaty Flavor -)

  • 정석근;이승규;김동훈;함준상
    • Journal of Dairy Science and Biotechnology
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    • 제26권1호
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    • pp.21-26
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    • 2008
  • Since goat milk infant formula has been increased, it is expected that goat milk consumption would be increased. This review summarizes the characteristics of goat milk especially, milk fat, somatic cell count, and goaty flavor. Average milk fat content for one year of twelve goat milk farms was 3.6%, but $2.9{\sim}3.1%$ in summer, which means summer goat milk could not meet the 'Processing and Ingredient Standard for Animal Products'. More than 3.2% for goat milk fat content in 'Processing and Ingredient Standard for Animal Products' should be amended. In addition to, hygienic standard for goat milk should be newly established because goat milk has naturally higher somatic cell count with noninfectious factors. It is thought that 6-trans nonenal and some branched fatty acids are responsible for the goaty flavor. It is necessary to minimize goaty flavor from farm to table because goaty flavor is the most important factor for the promotion of goat milk industry.

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The potential of mesenchymal stem cells derived from amniotic membrane and amniotic fluid for neuronal regenerative therapy

  • Kim, Eun Young;Lee, Kyung-Bon;Kim, Min Kyu
    • BMB Reports
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    • 제47권3호
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    • pp.135-140
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    • 2014
  • The mesenchymal stem cells (MSCs), which are derived from the mesoderm, are considered as a readily available source for tissue engineering. They have multipotent differentiation capacity and can be differentiated into various cell types. Many studies have demonstrated that the MSCs identified from amniotic membrane (AM-MSCs) and amniotic fluid (AF-MSCs) are shows advantages for many reasons, including the possibility of noninvasive isolation, multipotency, self-renewal, low immunogenicity, anti-inflammatory and nontumorigenicity properties, and minimal ethical problem. The AF-MSCs and AM-MSCs may be appropriate sources of mesenchymal stem cells for regenerative medicine, as an alternative to embryonic stem cells (ESCs). Recently, regenerative treatments such as tissue engineering and cell transplantation have shown potential in clinical applications for degenerative diseases. Therefore, amnion and MSCs derived from amnion can be applied to cell therapy in neuro-degeneration diseases. In this review, we will describe the potential of AM-MSCs and AF-MSCs, with particular focus on cures for neuronal degenerative diseases.

Application of the modified handmade cloning technique to pigs

  • Lee, Eun Ji;Ji, Kuk Bin;Lee, Ji Hye;Oh, Hyun Ju;Kil, Tae Young;Kim, Min Kyu
    • Journal of Animal Science and Technology
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    • 제63권2호
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    • pp.281-294
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    • 2021
  • Although somatic cell nuclear transfer (SCNT) is frequently employed to produce cloned animals in laboratories, this technique is expensive and inefficient. Therefore, the handmade cloning (HMC) technique has been suggested to simplify and advance the cloning process, however, HMC wastes many oocytes and leads to mitochondrial heteroplasmy. To solve these problems, we propose a modified handmade cloning (mHMC) technique that uses simple laboratory equipment, i.e., a Pasteur pipette and an alcohol lamp, applying it to porcine embryo cloning. To validate the application of mHMC to pig cloning, embryos produced through SCNT and mHMC are compared using multiple methods, such as enucleation efficiency, oxidative stress, embryo developmental competence, and gene expression. The results show no significant differences between techniques except in the enucleation efficiency. The 8-cell and 16-cell embryo developmental competence and Oct4 expression levels exhibit significant differences. However, the blastocyst rate is not significantly different between mHMC and SCNT. This study verifies that cloned embryos derived from the two techniques exhibit similar generation and developmental competence. Thus, we suggest that mHMC could replace SCNT for simpler and cheaper porcine cloning.

Effects of Capsaicin on Adipogenic Differentiation in Bovine Bone Marrow Mesenchymal Stem Cell

  • Jeong, Jin Young;Suresh, Sekar;Park, Mi Na;Jang, Mi;Park, Sungkwon;Gobianand, Kuppannan;You, Seungkwon;Yeon, Sung-Heom;Lee, Hyun-Jeong
    • Asian-Australasian Journal of Animal Sciences
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    • 제27권12호
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    • pp.1783-1793
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    • 2014
  • Capsaicin is a major constituent of hot chili peppers that influences lipid metabolism in animals. In this study, we explored the effects of capsaicin on adipogenic differentiation of bovine bone marrow mesenchymal stem cells (BMSCs) in a dose- and time-dependent manner. The BMSCs were treated with various concentrations of capsaicin (0, 0.1, 1, 5, and $10{\mu}M$) for 2, 4, and 6 days. Capsaicin suppressed fat deposition significantly during adipogenic differentiation. Peroxisome proliferator-activated receptor gamma, cytosine-cytosine-adenosine-adenosine-thymidine/enhancer binding protein alpha, fatty acid binding protein 4, and stearoyl-CoA desaturase expression decreased after capsaicin treatment. We showed that the number of apoptotic cells increased in dose- and time-dependent manners. Furthermore, we found that capsaicin increased the expression levels of apoptotic genes, such as B-cell lymphoma 2-associated X protein and caspase 3. Overall, capsaicin inhibits fat deposition by triggering apoptosis.

Interspecies Somatic Cell Nuclear Transfer Technique for Researching Dog Cloning and Embryonic Stem Cells

  • Sugimura, Satoshi;Sato, Eimei
    • Asian-Australasian Journal of Animal Sciences
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    • 제24권1호
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    • pp.1-8
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    • 2011
  • Large quantities of high-quality recipient oocytes with uniform cytoplasm are needed for research in the promising field of somatic cell nuclear transfer (SCNT) and embryonic stem cell research. In canines, however, it is difficult to obtain large quantities of oocytes because each donor produces a limited number of mature oocytes in vivo. Although in vitro maturation (IVM) is considered an alternative approach to oocyte production, this technique is still too rudimentary to be used for the production of highquality, uniform oocytes in large quantities. One technique for overcoming this difficulty is to use oocytes obtained from different species. This technique is known as interspecies SCNT (iSCNT). This review provides an overview of recent advances in canine - porcine interspecies SCNT.

An occurrence of mammary spindle cell carcinoma in a dog

  • Hong, Sunhwa;Lee, Hyun-A;Kim, Dong-Woo;Kim, Tae-Wan;Kim, Okjin
    • 한국동물위생학회지
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    • 제37권4호
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    • pp.313-317
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    • 2014
  • A bitch was presented for investigation of the mass in left 5th mammary gland. The partial mastectomy was performed and submitted for the histopathological diagnosis. The mammary mass was firm and white colored. The cut surface was separated with several lobules and developed vessels. The central area of the mass formed the cavity filled with inflammatory exudates. The dominant component of the tumor was the bundles of spindle-shaped cells. Some tumor cells possessed atypical nuclei and were arranged in a solid nest. Cysts were microscopically composed of hemorrhage, necrosis, and exudates, partially surrounded by tumor cells and granulation tissues. Histopathologically, the mammary mass revealed spindle cell carcinoma. The bitch made a complete recovery following the mastectomy. This case was a rare mammary spindle cell carcinoma in a dog.

미성숙 돼지 정소에서 pregnancy-associated plasma protein-A의 발현의 세포학적 분석 (Cytological analysis of pregnancy-associated plasma protein-A expression in porcine neonatal testis)

  • 김지윤;오건봉;변승준;옥선아;이휘철;황성수;박상현;하우태;우제석;송혁
    • 한국수정란이식학회지
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    • 제33권3호
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    • pp.177-183
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    • 2018
  • 생체 조직 내에서 표지인자의 발견은 해당 세포의 특성과 기능을 이해하는 데 매우 중요하다. 이전의 연구에서 본 연구진은 돼지정원줄기세포에서 특이적으로 IGFBP 3가 발현되어 표지인자로의 가능성을 보고한 바 있다. 본 연구에서는 IGFBP 3이외의 다른 family member가 정원줄기세포에서 특이적으로 발현하는 지와, 이의 발현을 조절하는 PAPP-A의 조직학적인 측면에서의 발현 양상을 5일령 돼지 정소에서 확인하였다. 그 결과 IGFBP 1, 2, 3, 4, 6의 발현은 정소 전체에서 발현되는 수준보다 돼지 정원줄기세포에서 더 높은 수준에서 발현하고 있음을 확인하였다. PAPP-A는 sertoli cell에서 특이적으로 발현하며, 정원줄기세포에서는 발현하지 않는 것을 PGP9.5와의 동시-조직염색으로 확인하였다. 이러한 결과는 Sertoli cell에서 발현하는 PAPP-A 단백질은 미성숙 돼지 정소에서 IGFBP family를 통해 정소 세포의 발달과 분화를 조절할 것으로 판단된다.