• 제목/요약/키워드: Mammalian cell

검색결과 734건 처리시간 0.031초

Peri-implanting 단계의 돼지배아 EST 연구 (The EST Study of the Peri-implanting Porcine Embryos)

  • 곽인석
    • 생명과학회지
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    • 제19권5호
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    • pp.587-592
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    • 2009
  • 임신초기의 배아는 peri-implantation 단계에서 매우 극적인 형태학적변화가 일어나는데, 이는 임신 인식 인자로 작용하는 배아에서 제공된 signal(s)에 의해서 시작되어지며, 나아가서 모성자궁과 배아의 상호 신호전달이 임신의 시작과 유지에 필수적인 인자로 작용한다. 배아형태의 급격한 리모델링에 관련된 세포학적, 생화학적, 유전학적 연구를 위하여, 또한 자궁과 배아의 상호 신호전달에 관여하는 잠재적 유전자 군을 발굴하기 위하여, peri-implantation 시기의 돼지배아를 이용하여 expresses sequences tag (EST) 분석을 실행하였다. 돼지배아 EST 분석으로 임신초기 특히 전 착상 단계에서 발현되는 유전자들의 카탈로그(Transcriptome)를 작성하였다. 그중에서 6개의 clone을 선택하여 그 발현 양식을 배아 및 자궁 등에서 관찰한 결과, 각각의 유전자들은 조직, 세포 유형 및 임신 시기에 따른 특이적인 발현 현상을 나타내었다. 본 연구결과는, 배아와 자궁 내막에서의 유전자 발현이 임신 시기에 따라서 다이내믹한 상호 조절 작용을 하고 있음을 나타낸다. 이는 전 착상단계의 모성자궁에서 배아와 자궁 내막의 상호 신호전달이 전 착상 단계의 배아의 급격한 형태학적 변화를 가능하게하고 또한 착상에 필요한 적절한 자궁 내부 환경을 제공하고 있음을 보여준다.

Implantation in Ruminants: Changes in Pre-Implantation, Maternal Recognition of Pregnancy, Control of Attachment and Invasion - Review -

  • Nagaoka, K.;Yamaguchi, H.;Aida, H.;Yoshioka, K.;Takahashi, M.;Christenson, R.K.;Imakawa, K.;Sakai, S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제13권6호
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    • pp.845-855
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    • 2000
  • As high as 50% of pregnancies are known to fail and the majority of such losses occur during the peri-implantation period. For the establishment of pregnancy in mammalian species, therefore, implantation of the conceptus to the maternal endometrium must be completed successfully. Physiological events associated with implantation differ among mammals. In ruminant ungulates, an elongation of the trophohlast in early conceptus development is required before the attachment of the conceptus to the uterine endometrium. Moreover, implantation sites are restricted to each uterine caruncula where tissue remodeling, feto-maternal cell fusion and placentation take place in a coordinated manner. These unique events occur under strict conditions and are regulated by numerous factors from the uterine endometrium and trophoblast in a spatial manner. Interferon-tau (IFN-${\tau}$), a conceptus-derived anti-Iuteolytic factor, which rescues corpus luteum from its regression in ruminants, is particularly apt to play an important role as a local regulator in coordination with other factors, such as TGF-${\beta}$, Cox-2 and MMPs at the attachment and placentation sites.

Antioxidative Effect of Astaxanthin on In Vitro Development of Porcine IVF Embryos

  • Jeon, Gi-Jun;Jang, Hyun-Yong;Cheong, Hee-Tae;Kim, Jong-Taek;Park, Choon-Keun;Chu, Duong-Xuan;Lee, Hak-Kyu;Yang, Boo-Keun
    • Reproductive and Developmental Biology
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    • 제31권1호
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    • pp.49-53
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    • 2007
  • Astaxanthin is a kind of carotenoid compounds, having a antioxidant and anti-inflammatory activities. The antioxidative mechanism by which carotenoid scavenge free radicals has been clearly elucidated, but has not tried for the development of mammalian preimplantation embryo. This study was conducted to investigate the antioxidative effect of astaxanthin on in vitro development of porcine in vitro fertilized embryos. Porcine embryos derrived from in vitro fertilization (IVF) were cultured in 5% $CO_{2} in air at $38.5^{\circ}C$ in PZM-3 medium supplemented with different dosages of astaxanthin ($0,\;1,\;5\;and\;10{\mu}M$) and taurine (0, 1, 2.5 and 5 mM) as a positive control, and execute to compare the effects of various antioxidants such as taurine, melatonin and asculatin on in vitro development. The proportions of embryos developed to the blastocyst stage were increased when $1\;and\;5\;{\mu}M$ of astaxanthin (26.6 and 23.4%, respectively) and 1 and 2.5 mM taurine (25.8 and 26.4%, Respectively) were supplemented, compared to controls (p<0.05). Also, various antioxidant-treated groups were significantly higher rates of blastocysts (astaxanthin, 27.4%; taurine, 29.1%; melatonin, 26.8%; aesculetin, 27.9%, respectively) than control (18.8%). There was no difference in mean cell number of blastocysts between antioxidants and control. This result indicates that astaxanthin has an antioxidant feature when porcine IVF embryos were cultured in vitro.

생쥐의 생식소 분화과정중 난소내 Gonadotropin-Releasing Hormone 유전자의 발현 (Expression of Gonadotropin-Releasing Hormone Gene in Mouse Fetal Ovary during Gonad Differentiation)

  • 윤성희
    • 한국발생생물학회지:발생과생식
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    • 제1권2호
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    • pp.189-202
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    • 1997
  • The hypothalamic peptide GnRH plays a central role in the regulation of the mammalian reproductive axis. Recent studies suggested that GnRH stimulates or inhibits the ovarian steroidogenesis and gametogenesis directly. Our previous report indicated that GnRH gene is expressed in adult rat ovary as well as in hypothalamus and that the expressed GnRH may induce the follicular atresia and apoptosis of ovarian granulosa cells in rat. Therfore, we studied whether GnRH gene is expressed in the mouse fetal ovary, when the germ cells are degenerating by apoptosis during gonad diffeerentiation. Mouse fetal gonads were obtained on the 12, 15,18 and 20th day of gestation from the mother mice superovulated (10 IU PMSG and 10 IU hCG) and mated. The morphological changes of fetal ovaries were examined histochemically by hematoxylin-eosin staining. The fetal sex was confirmed by PCR methods for sexing. RT-PCR methods were used to examine the expression of GnRH gene and the sex steroid hormones were determined by conventional radioimmunoassays. The levels of estradiol (E) and progesterone (P) were increaseduntil 18th day of gestation and then E was decreased just before parturition. The morphological changes of fetal gonadal tissue sections showed the ovarian development and coincided with the result of PCR analysis for sexing using ovary- or testis- specific oligonucleotide primers. Immunoreactive GnRH in placenta was decreased gradually until the end of gestation but fetal brain and ovarian GnRH were increased. The level of GnRH gene expression was increased during fetal ovarian development from 12 till 18th day and decreased suddenly on 20th day just before birth. From these results, it is suggested that ovarian GnRh may play a regulatory role on the germ cell differentiation of fetal ovary.

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포유동물 난자의 유성 및 무성 발생과정 동안 핵 및 세포질의 변화 (Nuclear and Cytoplasmic Dynamics in Mammalian Oocytes during Sexual and Asexual Developments)

  • Kim, Nam-Hyung
    • 한국발생생물학회지:발생과생식
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    • 제4권1호
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    • pp.7-12
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    • 2000
  • 수정에 의한 배 발생은 정자가 난자 내로 침입하여 정자와 난자의 반수체 핵질이 융합되고 이어 유사분열로 이어지는 과정에서 시작된다. 하지만 수정 및 초기 배 발생 동안 자웅 핵질과 난 세포질 구성 요소 상호간의 작용기전에 관해서는 명확히 알려져 있지 않은 부분이 많다. 수정보조기법인 세포질 내 정자 직접 주입법의 개발은 남성불임치료에 혁신적인 기술로 자리잡고 있을 뿐만 아니라 포유동물의 수정과정을 이해하는데 많은 도움을 주고 있다. 핵치환에 의한 복제동물 생산기법도 분화된 핵이 난 세포질 내에서 재 분화 (reprogramming)하여 발생하는 유일한 과정으로 세포질 구성요소들의 상호작용과 발생 조절 기능을 이해하는데 도움을 준다. 최근 몇 년간 돼지 난자 세포질에 정자 및 원형정자 직접주입, 세포질 이식, 세포질 융합 및 핵치환 한 후 난자의 발생과정을 간접 면역형광 분석법과 주사 전자현미경으로 조사하였다. 이러한 연구를 통해 체외수정, 세포질 이식 및 정자직접 주입법 등과 같은 임상치료기술 과 핵치환에 의한 복제동물생산 기법의 개선에 필요한 기초자료를 얻을 수 있었고, 포유동물 난자의 후생적 발생과정 (epigenesis)에 관해 공부할 수 있었다.

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에이코사펜타인산이 돼지난포란의 체외 성숙에 미치는 영향 (Effects of Eicosapentaenoic Acid during In Vitro Maturation of Porcine Oocytes: Hormone Synthesis and Embryonic Developmental Potential)

  • 김강식;박흠대
    • 한국동물생명공학회지
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    • 제34권3호
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    • pp.222-231
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    • 2019
  • Among fatty acid families, the polyunsaturated fatty acids were demonstrated to be mediators in various reproductive processes as precursor of steroid hormone (via cholesterol) and prostaglandins (via arachidonic acid), and in the last decade, major research was focused on the effects of omega-6 and especially omega-3 fatty acid. Eicosapentaenoic acid, the longest members of omega-3 fatty acid family, can be produced by a series of desaturation and elongation reactions from shorter member such as α-Linolenic acid. However, very few studies have provided detailed descriptions of Eicosapentaenoic acid effects and mechanisms of action in mammalian oocytes. The purpose of this study was to evaluate the effect of Eicosapentaenoic acid supplementation on in vitro maturation and developmental potential of porcine oocytes. Various concentrations of Eicosapentaenoic acid was added into in vitro maturation medium, and we evaluated the degree of cumulus expansion, nuclear maturation rate, blastocysts quality, and levels of prostaglandin E2, 17β-estradiol, progesterone in the spent medium. High doses (100 μM) of Eicosapentaenoic acid supplementation significantly inhibited cumulus expansion and oocyte nuclear maturation, and prostaglandin E2 synthesis also significantly decreased compared with other groups (p < 0.05). Supplementation of 50 μM Eicosapentaenoic acid showed higher quality blastocysts in terms of high cell numbers and low apoptosis when compared with other groups (p < 0.05), and synthesis ratio of E2/P4 also significantly increased compared with control group (p < 0.05). However, Supplementation of 100 μM Eicosapentaenoic acid showed high apoptosis when compared with other groups (p < 0.05), and synthesis ratio of 17β-estradiol/progesterone also significantly decreased compared with control group (p < 0.05). Our results indicated that supplementation with appropriate levels of Eicosapentaenoic acid beneficially affects the change of hormone synthesis for controlling oocyte maturation, leading to improved embryo quality. However, high doses of Eicosapentaenoic acid treatment results in detrimental effects.

십전대보탕이 신경교세포의 산화적 손상에 미치는 영향 (Effects of Sibjeondaebo-Tang on Oxidative Stress of C6 Glial Cells)

  • 류지용;윤종민;조광호;문병순
    • 동의생리병리학회지
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    • 제18권4호
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    • pp.1120-1128
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    • 2004
  • Zinc ion has both essential and toxic effects on mammalian cells. The results demonstrated that the ability of zinc to act as an inducer of apoptosis in C6 glial cells. Incubation with 0.2 mM ZnCl₂ caused cell death that was characterized as apoptosis by internucleosomal DNA fragmentation, formation of apoptotic bodies, nuclear fragmentation and breakdown of the mitochondrial membrane potential. ZnCl₂-induced apoptosis of C6 glial cells was prevented by the addition of Sibjeondaebo-Tang and antioxidants including reduced glutathione, N-acetyl-L-cysteine. We further confirmed that ZnCl₂ decreased the intracellular levels of GSH as well as generation of H₂O₂ in C6 glial cells. In 2D-electrophoresis, computer-assisted comparative analysis of the respective silver stained spot patterns revealed 3 groups with strongly decreased intensity by ZnCl₂. Whereas, 3 groups with increasing intensity were recovered by Sibjeondaebo-Tang. These results suggest that Sibjeondaebo-Tang may function as an antioxidant against free radicals and be applicable to the treatment of brain cells against oxidative stress.

남생이 위장관 및 췌장 내분비세포에 관한 면역조직화학적 연구 (An Immunohistochemical study of the gastro-entero-pancreatic endocrine cells in the alimentary tract and the pancreas of the fresh water turtle, Geoclemys reevesii)

  • 김종범;이재현
    • 대한수의학회지
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    • 제32권3호
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    • pp.321-331
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    • 1992
  • The regional distribution and relative frequency of gastrointestinal endocrine cells were studied immunohistochemically in the gastrointestinal mucosa and pancreas of the fresh water turtle. Ten kinds of endocrine cells were identified in the gastrointestinal tract. Cholecystokinin-8-, bovine pancreatic polypetide-and glucagon-immunoreactive cells were seen throughout the gastrointestinal tract, also among them cholecystokinin-8-immunoreactive cells were most predominant in segment III. Although gastrin- and gastrin/cholecystokinin-immunoreactive cells were found from segment III to VI and X, respectively, they were numerous in segment III. Somatostatin-immunoreactive cells were observed from segment I to VII. 5-hydroxytryptamine- immunoreactive cells were detected in segment I, III, VIII, IX and X. Human pancreatic polypeptide-immunoreactive cells were demonstrated in segment V, VI, VIII, IX and X. Insulin-immunoreactive cells were found from segment III to X except for segment VIII, but rare in segment VII. Neurotensin-immunoreactive cells were found to be restricted to segment VIII, IX and X. No porcine chromogranin-, substance P- and bombesin-immunoreactive cells were detected throughout the gastrointestinal tract of the fresh water turtle. Although typical mammalian pancreatic islets encapsulated by connective tissue were not present in this species, five kinds of endocrine cells-glucagon, insulin, somatostatin, bovine pancreatic polypeptide and 5-hydroxytryptamine-were found in forming small or large groups and scattered in the exocrine gland region. However porcine chromogranin- and motilin-immunoreactive cells could not be demonstrated in the pancreas.

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한약이 임신중 태아에 미치는 영향(II) -한약이 돌연변이원성과 염색체이상에 미치는 효과- (Studies on the Effects of Herbal medicines on the Fetus during Pregnancy (II) - Mutagenesis and chromosomal aberration of herbal medicines -)

  • 김동현;김남재;장준복;송병기
    • 대한한의학회지
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    • 제20권2호
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    • pp.121-127
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    • 1999
  • Oriental herbal medicines were examined for mutagenicity in the reverse mutation test on Salmonella typimurium T A98/100 and chromosomal aberration test on cultured mammalian cells (Chinese hamster cell lines). The reverse mutation test was performed by a plate incorporation method with and without a metabolic activation system (S9 mix). The tested herbal medicines did not significantly increase revertible colonies on any of the test strains with and without a metabolic activation system (S9 mix) at concentrations of 1 mg/ml. In the chromosomal aberration test, most tested herbal medicines did not significantly increase the number of aberrant cells on any of the test strains with a metabolic activation system (S9 mix) at concentrations of 1 mg/ml, compared with the vehicle control. However. Ansu Semen significantly increased the number of aberrant cells without a metabolic activation system (S9 mix). Paeoniae Radix. Hoelen, Aurantii nobilis Pericarpium, Cnidii Rhizoma, Angeliacae gigantis Radix, Perillae Herba and Moutan Cortex Radicis slightly increased revertible colonies on any of the test strains with a metabolic activation system (S9 mix), These results indicate that most herbal medicines might be carefully used in obstetrics and gynecology, although they do not have the potent mutagenic potential under the present test conditions.

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뉴켓슬병 바이러스에 대한 단크론성 항체생산 및 진단에 이용 (Production of Monoclonal Antibody to Newcastle Disease Virus and its Diagnostic Use)

  • Chung Ok Choi;Chung Gil Lee;Sung Man Cho;Jin Soo Na;Soo Hwan An;Joon Hun Kwon
    • 한국가금학회지
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    • 제15권3호
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    • pp.207-210
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    • 1988
  • 뉴켓슬병바이러스(NDV)인 LaSota 주를 SPF 발육난의 요막강내에 증식시켜 순수 정제한 것을 BALB/c 흰쥐에 면역시킨 후 추출한 비강세포와 흰쥐 골수암세포와의 융합방법에 의하여 NDV에 특이하게 작용하는 단크론성항체(MCA)를 생산하는 3주의 Hybridoma틀 작성하였다. 이 3주의 MCA는 모두 IgG형에 속하였으며 흰쥐 복강 내에 접종하여 생산된 복수항체의 항체가는 간접형광항체법으로 $10^3$-$10^6$에 달하였고 약독 및 강독 NDV에 모두 동일한 수준으로 작용하였다. 중화능은 인정되지 않았고 3주중 1주만이 별구응집 억제능을 약하게 나타냈다. 이 MCA를 이용하여 간접형광항체법으로 인공 감염시킨 닭에서 NDV항원 검출을 시도한 결과 기관점막을 비롯한 각종 장기의 도말표본에서 접종 3일 후부터 뚜렷한 검출이 가능하였다.

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