• 제목/요약/키워드: follicles

검색결과 832건 처리시간 0.024초

The Effect of Fibroblast Co-culture on In Vitro Maturation of Mouse Preantral Follicles

  • Kim, Chung-Hoon;Cheon, Yong-Pil;Lee, You-Jeong;Lee, Kyung-Hee;Kim, Sung-Hoon;Chae, Hee-Dong;Kang, Byung-Moon
    • 한국발생생물학회지:발생과생식
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    • 제17권3호
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    • pp.269-274
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    • 2013
  • This study was performed to evaluate the effects of fibroblast co-culture on in vitro maturation (IVM) of prepubertal mouse preantral follicles. The intact preantral follicles were obtained from the ovaries of 12-14 day old mice and these were cultured individually in ${\alpha}$-minimal essential medium (${\alpha}$-MEM) supplemented with 5% fetal bovine serum (FBS), $100mIU/m{\ell}$ recombinant follicle stimulating hormone (rFSH), 1% insulin-transferrin-selenium, $100{\mu}g/ml$ penicillin and $50{\mu}g/m{\ell}$ streptomycin as base medium for 12 days. A total of 200 follicles were cultured in base medium co-cultured with mouse embryonic fibroblast (MEF) (MEF group) (n=100) or only base medium as control group (n=100). Survival rate of follicles on day 12 of culture were significantly higher in the MEF group of 90.0%, compared with 77.0% of the control group (p=0.021). Follicle diameters on day 6 and 8 of the culture period were significantly larger in the MEF group than those in the control group (p=0.021, p=0.007, respectively). Estradiol levels in culture media on day 4, 6, 8, 10 and 12 of the culture period were significantly higher in the MEF group (p=0.043, p=0.021, p=0.006, p<0.001 and p=0.008, retrospectively). Our data suggest that MEF cell co-culture on IVM of mouse preantral follicle increases survival rate and promotes follicular growth and steroid production.

방사성동위원소옥소(放射性同位元素沃素)($^{131}I$)가 "마우스"의 주요장기(主要臟器)의 병리조직상(病理組織像)에 미치는 영향(影響)에 관(關)한 연구(硏究) (Histopathological Studies of Mice after Administration of Radioactive Iodine($^{131}I$))

  • 노재성;이문호
    • 대한핵의학회지
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    • 제1권2호
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    • pp.85-97
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    • 1967
  • Histopathological changes of various organs of the mice after intra-peritoneal injections of radioactive iodine ($^{131}I$) were experimentally observed. Sixity healthy female mice, weighing average 25 gm, devided into 6 groups, were used. The various doses of $^{131}I$ were injected intraperitoneally at different intervals. The histopathological changes after these treatments were observed in organs such as thyroids, parathyroids, livers, kidneys and gonads. Following were the results; 1) Thyroid: In the group A given $^{131}I$ with a single dose of $10{\mu}C$ per gm body weight, it was observed that the protoplasms of follicular epithelial cells were destroyed, the nuclei were expanded or dissoluted, showing pyknotic changes of nuclei and vacuolizations of protoplasms. In the group B given $^{131}I$ with a single dose of $5{\mu}C$ per gm body weight, hyperemias, hemorrhages and hyaline degenerations in the whole area were observed. In the group C given $^{131}I$ with 3 doses of $2.5{\mu}C$ per gm body weight every week, the thyroid parenchyms were destroyed and epithelial cells of varing size were observed in the fibrinous tissues. In the group D given $^{131}I$ with 6 doses of $0.5{\mu}C$ per gm body weight every week, some destroyed follicles and new borne follicles were observed. But the histopathological changes resemble the follicles of the normal thyroid gland. In the group E and F given $^{131}I$ with 8 and 10 doses of $0.2{\mu}C\;and\;0.01{\mu}C$ for each group per gm body weight every two days, both pyknotic changes of nuclei and cytoplasmic vacuolizations of the follicular epithelia, hypertrophies of follicles and abnormal irregular follicular structures were observed, and in the group F, lymphocytes appeared around the thyroid glands. 2) Parathyroid: In the group A, hyperemia, proliferations of connective tissues, karyorrhexes and vacuolizations were observed. In other experimental groups, no particular pathological change was observed. 3) Liver: The degnerative changes and acute or chronic inflammatory changes were observed in proportion to the amount of $^{131}I$ injected. Atrophies of the liver cells, dilatations of sinusoids, hyaline degenerations and necrotic pictures were observed. 4) Kidney: In the group A, congestions and infiltrations of mononuclear cells and granulocytes were observed around the cortical arteries, and in the group B, the degenerative changes of cortexes, and, in the group C and D, hydronephrotic changes were observed respectively, and hyaline degenerations were partially observed. 5) Gonad: In the group A, the follicles were degenerated. The ova in the follicles showed irregular figures. The changes in the group B were almost the same as in the group A, but the changes were mild. In the group C, the destructions of whole ova, the hypertrophies of ovarian follicular membranes and pyknotic changes of nuclei were observed. In the group D, the pathological changes were similar to that of group C, but mild in the grade. In the group E, almost none of ovarian follicular fluid was observed, and in the group F, the tissue pictures were almost similar to that of the normal group.

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In vitro growth of mouse preantral follicles: effect of animal age and stem cell factor/insulin-like growth factor supplementation

  • Jee, Byung Chul;Kim, Jee Hyun;Park, Da Hyun;Youm, Hyewon;Suh, Chang Suk;Kim, Seok Hyun
    • Clinical and Experimental Reproductive Medicine
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    • 제39권3호
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    • pp.107-113
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    • 2012
  • Objective: To determine whether animal age impacts in vitro preantral follicle growth. Effects of hCG, stem cell factor (SCF), and/or insulin-like growth factor (IGF) supplementation in growth medium were also investigated. Methods: Intact preantral follicles were mechanically isolated from fresh ovaries of BDF1 mice and cultured in growth medium for 9 to 11 days. Surviving follicles with antrum formation were transferred to maturation medium for 14 to 18 hours. Follicle survival, antrum formation, and retrieval of metaphase II (MII) oocytes were compared among three age categories (4-5, 7-8, and 10-11 week-old). By using 7- to 8-week-old mice, preantral follicles were cultured in growth medium supplemented with hCG (0, 5, or 10 mIU/mL), SCF (50 ng/mL), IGF-1 (50 ng/mL), and SCF+IGF-1. Results: Seven- to eight-week-old mice showed a higher follicle survival and antrum formation and produced more MII oocytes compared to other groups. In the 7- to 8-week-old mice, supplementation of 5 mIU/mL hCG significantly enhanced the antrum formation but the percentage of MII oocytes was similar to that of the control. Supplementation of SCF+IGF-1 did not enhance follicle survival or antrum formation but the percentage of MII oocytes increased modestly (39.1%) than in the control (28.6%, p>0.05, statistically not significant). Conclusion: Seven- to eight-week-old mice showed better outcomes in growth of preantral follicles in vitro than 4- to 5- or 10- to 11-week-old mice. Supplementation of hCG enhanced antrum formation and supplementation of SCF+IGF-1 yielded more mature oocytes; hence, these should be considered in the growth of preantral follicles in vitro.

Induction of Fas-Mediated Apoptosis by Interferon-g is Dependent on Granulosa Cell Differentiation and Follicular Maturation in the Rat Ovary

  • Lee, Hye-Jeong;Kim, Ji Young;Park, Ji Eun;Yoon, Yong-Dal;Tsang, Benjamin K.;Kim, Jong-Min
    • 한국발생생물학회지:발생과생식
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    • 제20권4호
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    • pp.315-329
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    • 2016
  • Fas ligand (FasL) and its receptor Fas have been implicated in granulosa cell apoptosis during follicular atresia. Although interferon-gamma (IFN-${\gamma}$) is believed to be involved in the regulation Fas expression in differentiated granulosa or granulosa-luteal cells, the expression of this cytokine and its role in the regulation of the granulosa cell Fas/FasL system and apoptosis during follicular maturation have not been thoroughly investigated. In the present study, we have examined the presence of IFN-${\gamma}$ in ovarian follicles at different stage of development by immunohistochemistry and related their relative intensities with follicular expression of Fas and FasL, and with differences in granulosa cell sensitivity to Fas activation by exogenous agonistic Anti-Fas monoclonal antibody (Fas mAb). Although IFN-${\gamma}$ immunostaining was detectable in oocyte and granulosa cells in antral follicles, most intense immunoreactivity for the cytokine was observed in these cells of preantral follicles. Intense immunoreactivity for IFN-${\gamma}$ was most evident in granulosa cells of atretic early antral follicles where increased Fas and FasL expression and apoptosis were also observed. Whereas low concentrations of IFN-${\gamma}$ (10-100 U/mL) significantly increased Fas expression in undifferentiated granulosa cells (from preantral or very early antral follicles) in vitro, very higher concentrations (${\geq}1,000U/mL$) were required to up-regulate of Fas in differentiated cells isolated from eCG-primed (antral) follicles. Addition of agonistic Fas mAb to cultures of granulosa cells at the two stages of differentiation and pretreated with IFN-${\gamma}$ (100 U/mL) elicited morphological and biochemical apoptotic features which were more prominent in cells not previously exposed to the gonadotropin in vivo. These findings suggested that IFN-${\gamma}$ is an important physiologic intra-ovarian regulator of follicular atresia and plays a pivotal role in regulation of expression of Fas receptor and subsequent apoptotic response in undifferentiated (or poorly differentiated) granulosa cells at an early (penultimate) stage of follicular development.

생쥐 Preantral Follicles의 체외성장 및 성숙에 있어서 Gonadotrophins의 역할 (Effects of Gonadotrophins on In Vitro Growth and Maturation of Mouse Preantral Follicles)

  • 김동훈;지희준;강희규;한성원;이훈택;정길생;이호준
    • 한국가축번식학회지
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    • 제23권1호
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    • pp.53-61
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    • 1999
  • 본 연구는 생쥐 preantral follicle 의 체외성장 및 성숙에 있어서 gonadotrophins 인 FSH 와 LH 의 효과를 조사하기 위하여 실시하였으며, 그 결과는 다음과 같다. 1. FSH 첨가군들은 대조군에 비하여 유의하게 높은 생존율과 성숙율을 나타냈으며, 100$m\ell$D/$m\ell$ 의 FSH 농도가 preantral follicle 의 체외배양에 적정한 농도인 것으로 나타났다. 2. HMG 첨가군은 FSH 첨가군보다, 통계적 유의차는 인정되지 않았지만, 높은 생존율과 성숙율을 나타냈다. 3. FSH 와 LH 의 첨가비율이 100$m\ell$U/$m\ell$ 대 10$m\ell$D/$m\ell$(10:1) 에서 가장 높은 생존율과 성숙율을 나타냈다. 4. FSH 혹은 HMG 첨가시, 정상적인 oestradiol 과 progesterone 분비양상을 나타냈으며, HMG 첨가군에서 유의하게 높은 농도의 oestradiol 과 progesterone 을 분비하였다. 이상의 결과를 종합해 볼 때, gonadotrophins 은 preantral follicle 의 체외성장 및 성숙뿐만 아니라 steroidogenesis 에서 중요한 역할을 수행한다는 것을 확인할 수 있었다.

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경남지방의 도태우에 불임과 관련된 난소의 형태학적 관찰 1. 난포와 황체의 출현에 대하여 (Morphological Observations of Ovaries in Relation to Infertility in Slaughtered Cows in Kyungnam Province 1. Appearance of follicles and corpus luteums in cow ovaries)

  • 양재훈;표병민;서득록;고필옥;강정부;김종섭;곽수동
    • 한국임상수의학회지
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    • 제19권2호
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    • pp.147-152
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    • 2002
  • Ovaries from total 192 slaughtered cows, 154 Korean native cows and 38 dairy cows were collected during the slaughtering process in Kimhae, Changyoung and Yangsan abattoirs in Kyungnam province from January 2001 to January 2002. Rates of pregnant and non-pregnant and ovarian findings were invested. Rates of pregnant cows in 192 slaughtered cows were 12.5% (24 cows) and in difference of cow breeds, 11.0% (17 cows) in 154 Korean native cows and 18.4% (7 cows) in 38 dairy cows from total 192 cows, respectively. Ages of fetuses in pregnant Korean native cows were mostly less than 4 months and ages of fetuses in dairy cows were mostly about 7-8 months. Cows which each diameter of follicles and corpus luteums in same cow was more than 5-6 mm in diameter were 69.8% (134 cows) in total 192 slaughtered cows and in difference of cow breeds, 64.7% (11 cows) in 17 Korean native cows and 57.1% (4 cows) in 7 dairy cows. Mean diameter of foliicles and corpus luteums in Korean native cows are 13.7$\pm$5.6$\times$ 11.2$\pm$4.6mm and 17.5$\pm$4.6$\times$14.6$\pm$4.0 mm in non-pregnat cows, and are 11.0$\pm$4.8$\times$9.1 $\pm$ 2.6mm and 21.2$\pm$2.9$\times$18.3$\pm$ 2.7 mm in pregnant cows, respectively. Mean diameter of follicles and corpus luteums in dairy cows are 15.8$\pm$7.1 $\times$ 14.3$\pm$ 6.0 mm and 20.3$\pm$5.9$\times$16.9$\pm$ 5.8 mm in non-pregnant cows, and are 10.1 $\pm$ 3.0$\times$9.2$\pm$2.3 mm and 23.0$\pm$ 1.7$\times$20.1 $\pm$ 1.3 mm in pregnant cows, respectivley. The above findings indicate that the co-appearance rate of follicles and corpus luteums in same cows are higher in both pregnant and non-pregnant cows. Compared in pregnant and non-pregnant cow ovaries, mean size of follicles are smaller in pregnant cows but size of corpus luteums are more larger in pregnant cows than in non-pregnant cows. Correlation of the follicle size (Y) and corpus luteum size (X) in same cows developed each other in inversive size. Those correlative formulas appeared to be Y = -0.2022X+17.175 in Korean native cows and Y= -0.5754 X+24.153 in dairy cows.

6 MeV LINAC 방사선 조사에 의한 생쥐 난소조직의 미세구조 변화 (Fine Structural Modification of Mouse Ovarian Tissue by Irradiation of 6 MeV LINAC Radiation)

  • 윤철호;문명진
    • Applied Microscopy
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    • 제33권2호
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    • pp.117-130
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    • 2003
  • 본 연구는 6 MeV LINAC에서 발생한 X-선을 생쥐 생체에 조사한 후, 방사선 조사선량에 따른 난소 조직의 미세구조적 변화를 광학 및 전자현미경으로 관찰하였다. X-선을 200 cGy의 선량을 조사한 후 적출한 난소조직에서 성장난포의 과립층세포는 핵이 응축되어 나타났고, 난자를 둘러싸고 있는 투명층은 정상 대조군에서보다 경계가 불규칙하게 관찰되었다. 400 cGy로 조사된 난소조직에서 과립층세포는 대부분 응축된 핵을 가지고 있었으며, 피폭된 과립층세포의 사멸에 의한 세포질의 단백질 변성이 확인되었다. 600 cGy의 선량을 조사한 난소의 조직표본에서 성장난포는 위축되어 있었으며, 난포동에는 백혈구가 침윤되어 있었고, 난포액은 정상난포와 달리 불균질한 물질로 채워져 있었다. 투과전자현미경으로 관찰한 결과, 200 cGy의 방사선을 조사한 성장난포에서 과립층세포는 정상난포와는 달리 불규칙한 모양을 하고 있었다. 과립세포 사이의 난포동에는 많은 세포성분의 부스러기가 산재하여 있었으며, 이들 주변에 호중구와 대식세포가 관찰되어, 이 시기에 이미 방사선 피폭에 의한 세포사의 기작이 진행되고 있음이 확인되었다. X-선을 600 cGy로 조사한 난소조직의 과립층세포는 세포의 형태나 핵의 변형이 현저하여, 일부 세포는 핵의 분절화가 진행되는 등, apoptosis의 전형적인 특징이 관찰되었다. 특히 성장난포의 난포동에서는 염증반응에 관여하는 호중구가 잘 관찰되었으며, 주변에 산재된 세포 파쇄물 및 apoptotic body와 함께 대식세포의 출현도 확인되었다.

배양중인 개구리 여포의 cAMP 흡수와 분해 (Uptake and Degradadon of cAMP by Frog Follides in vitro)

  • 권혁방;나철호;안련섭;김경진
    • 한국동물학회지
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    • 제34권2호
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    • pp.181-187
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    • 1991
  • 양서류 여포를 배양할 때 외부에서 cAMP를 첨가하면 호르몬에 의한 난자의 성숙이 억제된다는 많은 보고가 있었다. 그러나 실제 외부의 cAMP가 여포내로 들어간다는 보고는 아직 없다. 본 연구에서는 배양액내의 cAMP가 여포내로 침투해 들어가는 현상과 들어간 cAMP의 분해과정을 radioimmunoassay로 조사하였다. 개구리 여포를 배양하면서 배양액에 난자의 성숙을 억제하는 농도의cAMP(2.5 mM)를 첨가한 후 일정시간 간격으로 여포내 축척된 cAMP의 농도를 조사한 결과 2시간에서 이미 기본수준(여포당 약 3 p mole)의 수십배로 증가하였다.(여포당 90 p mole). cAMP를 포함한 배양액에서 6시간 배양 후 보통 배양액으로 옮겨 배양하면서 일정 시간마다 여포내 cAMP의 농도를 측정한 결과 6시간 내에 cAMP농도가 여포당 160 p mole에서 약 10 p mole로 급격히 낮아졌다. 그러나 18시간 후에도 기본 수준으로까지 내려가지는 않았다. 이러한 cAMP의 감소과정이 progesterone이나 isobuty methylxanthine (IBMX)에 크게 영향을 받지 않았다. 배양중인 여포를 forskolin(9 u m)으로 자극했을 때에는 기본 수준의 약 3배정도로 cAMP의 농도가 증가하였다. 본 결과는 배양액내의 cAMP가 여포내로 투과해 들어가고 이들은 곧 여포에 의해 분해된다는 것을 시사하고 있다.

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과배란유도 여성에서 우성난포의 성장속도 (Growth Rate of Dominant Follicles During Controlled Ovarian Hyperstimulation)

  • 황은주;구승엽;김용진;지병철;서창석;김석현;최영민;김정구;문신용
    • Clinical and Experimental Reproductive Medicine
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    • 제37권3호
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    • pp.253-259
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    • 2010
  • 목 적: 과배란유도 환자에서의 우성난포의 성장속도와 임상인자들과의 연관성을 평가하고자 하였다. 연구방법: 체외수정시술을 위한 과배란유도 313주기를 대상으로 혈중 호르몬 농도를 측정하고 과배란유도 주기 중 난포 직경의 변화를 초음파를 이용하여 연속적으로 측정하였다. 우성난포의 성장 속도를 계산하고, 임상지표와의 연관성을 분석하였다. 결 과: Gonadotropin releasing hormone 작용제와 길항제 주기 사이에 우성난포의 성장속도의 유의한 차이는 관찰되지 아니하였다. 우성난포의 성장속도와 환자의 연령, 체질량 지수, 생리 시작 3일의 follicle stimulating hormone, luteinizing hormone, 에스트라디올, 획득 난자의 수, 수정률 등의 임상지표는 유의한 연관성이 없었다. 결 론: 본 연구 결과는 우성난포의 성장속도는 난소 반응을 포함한 임상지표와 연관성이 없는 독립적인 변수임을 시사한다.

Immunohistological expression of cytochrome P450 1A2 (CYP1A2) in the ovarian follicles of prepubertal and pubertal rat

  • Hwang, Jong-Chan;Park, Byung-Joon;Kim, Hwan-Deuk;Baek, Su-Min;Lee, Seoung-Woo;Jeon, Ryoung-Hoon;Jang, Min;Bae, Seul-Gi;Yun, Sung-Ho;Park, Jin-Kyu;Kwon, Young-Sam;Kim, Seung-Joon;Lee, Won-Jae
    • 한국동물생명공학회지
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    • 제35권4호
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    • pp.329-337
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    • 2020
  • Cytochrome P450 1A2 (CYP1A2) is a member of the cytochrome P450 superfamily enzymes in mammals and plays a major role in metabolizing endogenous hormones in the liver. In recent days, CYP1A2 expression has been found in not only the liver but also other tissues including the pancreas and lung. However, little information is available regarding the expression of CYP1A2 in the ovary, in spite of the facts that the ovarian follicle growth and atresia are tightly associated with controls of endocrine hormonal networks. Therefore, the expression of CYP1A2 in the ovaries of prepubertal and pubertal rats was investigated to assess its expression pattern and puberty-related alteration. It was demonstrated that the expression level of CYP1A2 was significantly (p < 0.01) higher in the pubertal ovaries than prepubertal counterparts. At the ovarian follicle level in both groups, whereas CYP1A2 expression was less detectable in the primordial, primary and secondary follicles, the strongly positive expression of CYP1A2 was localized in the granulosa cell layers in the antral and pre-ovulatory follicles. However, the ratio of CYP1A2-positive ovarian follicle was significantly (p < 0.01) higher in the ovary of pubertal group (73.1 ± 3.1%) than prepubertal one (41.0 ± 10.5%). During the Immunofluorescence, expression of CYP1A2 was mainly localized in Fas-positive follicles, indicating the atretic follicles. In conclusion, these results suggested that CYP1A2 expression was mainly localized at the atretic follicular cells and affected by the onset of puberty. Further study is still necessary but we hypothesize that CYP1A2 expresses in the atretic follicles to metabolize residue of the reproductive hormones. These findings may have important implications for the fields of reproductive biology of animals.