• Title/Summary/Keyword: Immunocytochemical study

검색결과 106건 처리시간 0.242초

체외 배양된 닭 배반엽 세포에 대한 Retrovirus Vector를 이용한 유전자 전이 (Retrovirus Vector-Mediated Gene Transfer to the Chicken Blastodermal Cells Cultured In Vitro)

  • 박성준;구본철;권모선;최휘건;김태완
    • Reproductive and Developmental Biology
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    • 제34권3호
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    • pp.257-262
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    • 2010
  • The purpose of this study is to establish a basic culture system enabling in vitro culture of chicken blastodermal cells and to test the feasibility of retrovirus-mediated gene transfer to the cultured cells. The blastodermal cells were isolated from freshly laid eggs of stage X and cultured with or without STO feeder layer cells. Stem cell-like morphology was maintained after multiple passages and RT-PCR analysis proved expression of several stem cell specific genes. Immunocytochemical analysis using antibodies of anti-EMA-1 and anti-SSEA-1 also showed the feature of stem cells. Infection of the cultured blastodermal cells with LNCGW retrovirus vector resulted in successful transfer of foreign genes. The results of this study may be useful in establishing stem cell-mediated transgenic chicken production.

Immunocytochemical Localization of Glutamatergic Neurons in the Lateral Reticular Nucleus Projecting to Ansiform (Crus I and II) and Paramedian Cerebellar Lobules of the Rat

  • Lee, Hyun-Sook
    • Animal cells and systems
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    • 제2권1호
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    • pp.139-144
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    • 1998
  • I examined the projection of glutamatergic neurons in the lateral reticular nucleus into ansiform (crus l and ll) and paramedian lobules in the rat cerebellum using immunocytochemical methods with antiserum against glutamate combined with WGA-HRP histochemistry. The projections of glutamatergic neurons from the lateral reticular nucleus to crus l were most extensive in number among the three injection cases and the majority of projections originated at the dorsal to dorsomedial region of the ipsilateral magnocellular nucleus. Glutamate-immunoreactive cells projecting to crus ll were less extensive in number than those projecting to crus l and were mainly localized at the dorsomedial portion of the ipsilateral magnocellular nucleus. Double-labelled neurons projecting to crus l or crux ll were also located at ipsilateral subtrigeminal as well as contralateral magnocellular nuclei. Glutamatergic neurons projecting to paramedian lobules were moderate in number and mainly located at the dorsal area of the ipsilateral magnocellular nucleus. A few double-labelled cells were also found at ipsilateral subtrigeminal or contralateral magnocellular nuclei. The present study suggests that glutamate-immunoreactive neurons at the dorsal to dorsomedial magnocellular division of the lateral reticular nucleus may participate in the excitatory control of target neuronal activities at ipsilateral, posterior hemispheric lobules of the rat cerebellum.

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전액성 삼출액내 반응성 중피세포와 암종세포간의 감별진단에서 calretinin의 유용성 (Utility of Calretinin in Distinction between Benign Reactive Mesothelial and Carcinoma Cells in Serous Effusions)

  • 김병헌
    • 대한세포병리학회지
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    • 제12권2호
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    • pp.89-95
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    • 2001
  • The cytological distinction of carcinoma cells from reactive mesothelial cells in serous effusions nay be difficult or imposslble based on morphology alone, especially In specimens containing reactive mesothelial cells which form glandular or ball- or papillary-shaped conglomerates or which mimic malignant nuclear features. Calretinin is a newly reported immunocytochemical marker for mesothelial cells, which can potentially be utilized for facilitating this distinction. This study evaluated the usefulness of calretinin for the discrimination between reactive mesothelial and metastatic carcinoma cells in serous effusion. Immunocytochemical staining was undertaken on 33 benign reactive and 87 malignant serous effusion specimens with histologically confirmed diagnoses. The specimens including smears and cell blocks were stained with polyclonal antibody to calretinin by labelled streptavidin-biotin method. The positive expression of calretinin was noted In 32(97.0%) of 33 benign reactive effusions and 9(10.3%) of 87 malignant effusions. The sensitivity and specificity of the calretinin immunostaining for reactive mesothelial cells was 97.0% and 89.7%, respectively. In conclusion, calretinin is a useful marker for distinguishing between reactive mesothelial cells and carcinoma cells in serous effusions.

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한우(韓牛)의 위장관(胃腸管)에 존재(存在)하는 내분비세포(內分泌細胞)의 면역세포화학적(免疫細胞化學的) 연구(硏究) (Immunocytochemical study of the endocrine cels in the gastrointestinal tract of the Korean native cattle)

  • 조성환;키타무라 노부오
    • 대한수의학회지
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    • 제28권2호
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    • pp.251-259
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    • 1988
  • Regional distribution and relative frequency of endocrine cells in ten portions of the gastrointestinal tract of the Korean native cattle were observed by immunocytochemical methods using specific antisera against chromogranin, serotonin, somatostatin, glucagon, bovine pancreatic polypeptide(BPP), motilin, gastric inhibitory polypeptide(GIP), neurotensin, secretin, gastrin and substance P. The results observed are summarized as follows: In the abomasum, chromogranin-, serotonin-, somatostatin-, motilin-, glucagon-, gastrin-, and substance P-immunoreactive cells were found. Chromogranin-and serotonin-immunoreactive cells were more numerous in the fundic region than pyloric region. Somatostatin- and gastrinimmunoreactive cells were numerous in the pyloric region than in the fundic region. In the small intestine, chromogranin-, serotonin-, somatostatin-, glucagon-, BPP-, motilin-, gastrin-, GIP-, neurotensin-, secretin-, and substance P-immunoreactive cells were detected. Chromogranin-, somatostatin-, GIP- and secretin-immunoreactive cells were most numerous in the duodenum, while BPP-, motilin-, glucagon-, neurotensin- and substance P-immunoreactive cells were rarely seen in the small intestine. In the large intestine, chromogranin-, serotonin- and BPP-immunoreactive cells were widely distributed and most numerous in the rectum. Somatostatin-, glucagon- and substance P-immunoreactive cells were rarely seen in the large intestine.

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Localization of Cyclin B and Erk1/2 in Ovine Oocytes and MPF and MAPK Activities in Cytoplast and Karyoplast following Enucleation

  • Lee, Joon-Hee;Campbell, Keith H.S.
    • Reproductive and Developmental Biology
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    • 제35권4호
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    • pp.407-414
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    • 2011
  • The development of embryos reconstructed by somatic cell nuclear transfer (SCNT) is dependent upon numerous factors. Central to development is the quality and developmental competence of the recipient cytoplast and the type of the donor nucleus. Typically metaphase of the second meiotic division (MII) has become the cytoplast of choice. Production of a cytoplast requires removal of the recipient genetic material, however, it may remove proteins which are essential for development or reduce the levels of cytoplasmic proteins to influence subsequent reprogramming of the donor nucleus. In this study, enucleation at MII did not affect the activities of either MPF or MAPK kinases. Immunocytochemical staining showed that both Cyclin B1 (MPF) and Erk1/2 (MAPK) were associated with the meiotic spindle of AI/TI oocytes with little staining in the cytoplasm, however, at MII association of both proteins with the spindle had reduced and a greater degree of cytoplasmic distribution was observed. The analysis of oocyte proteins removed during enucleation is a difficult approach to the identification of factors which may be depleted in the cytoplast. This is primarily due to the large numbers of aspirated karyoplasts which would be required for the analysis.

인삼 종자의 저장단백질에 관한 면역 세포화학적 연구 - Tris 완충액 가용성 단백질 - (An Immunocytochemical Study on Storage Proteins of Ginseng Seed - Tris Buffer Soluble Protein -)

  • 김우갑
    • Applied Microscopy
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    • 제19권2호
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    • pp.74-84
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    • 1989
  • 인삼 종자의 배유조직에서 Tris 완충액 가용성 저장단백질을 추출한 후 전기 영동적 분석으로 분리하여 $SP_{1}$(MW=160,000)과 $SP_2$(MW=70,000)의 두가지 저장단백질을 정제하였다. 이 두가지 저장단백질을 항원으로 사용하여 토끼에 피하주사하여 항체를 얻었으며, 이 항체를 이용하여 면역 세포화학적 금입자표지법을 실시한 결과, $SP_1$$SP_2$ 모두 구형의 protein body내에 산재하여 있음을 확인되었으며, globoid에는 이러한 두가지 단백질중 어느 것도 함유되어 있지 않는 것으로 나타났다. 또한 각각의 protein body에 함유된 $SP_1$$SP_2$의 상대적 함량에는 서로 차이가 있음이 확인되었다.

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면역전현법(免疫電顯法)에 의한 마쇄(磨碎) 리그닌의 기원(起源)에 관한 연구(硏究) (Origin of Milled Wood Lignin : An Immunocytochemical Approaches in Combination with Transmission Electron Microscopy)

  • 김윤수;고홍범
    • Journal of the Korean Wood Science and Technology
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    • 제24권1호
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    • pp.68-74
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    • 1996
  • Present work was undertaken to investigate the origin of milled wood lignin(MWL) in the wood cell wall using immunocytochemical techniques, which can provide the information on the localization of specific antigens(MWL in the present study) to be examined. Spruce MWL dissolved in DMSO and emulsified with Freund adjuvant was injected directly into the mouse spleen. The animals were boostered at two-week intervals after the initial immunization. Blood samples were purified in standard procedures. The characteristics of antibodies against MWL were tested by indirect ELISA. Visualization of MWL was carried out using conventional indirect immunogold-labelling methods on the ultrathin sections of spruce wood. Immuno-TEM observations showed that the immunogold probes were selectively attached to secondary cell walls of spruce wood. The most intense labelling was frequently observed in the S2 layer. In contrast, gold labelling in the lignin-rich regions, such as middle lamella and cell corner was not found. The immuno-TEM provides an indication that spruce MWL originates from the S2 layer.

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Optimization of Human Embryonic Stem Cells into Differentiation of Dopaminergic Neurons in Vitro: II. Genetically Modified Human Embryonic Stem Cells Treated with RA/AA or b-FGF

  • 신현아;김은영;이영재;이금실;조황윤;박세필;임진호
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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    • pp.75-75
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    • 2003
  • Since the establishment of embryonic stem cell, pluripotency of the cells was known to allow differentiation of the cells into various cell types consisting whole body. Several protocols have been developed to induce expression of specific genes.. However, no precise protocol that will generate a single type of the cells from stem cells has been reported. In order to produce cells suitable for transplantion into brain of PD animal model, which arouse due to a progressive degeneration of dopaminergic neurons in midbrain, human embryonic stem cell (hESC, MB03) was transfected with cDNAs cording for tyrosine hydroxylase (TH). Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by the two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA/ascorbic acid (AA), embryoid bodies (EB, for 4days) derived from hES cells were exposed to RA (10$^{-6}$ M)/AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. By indirect immunocytochemical studies, proportion of cells expressing NF200 increased rapidly from 20% at 7 days to 70 % at 28 days in RA/AA-treated group, while those cells expressing NF160 decreased from 80% at 7 days to 10% at 28 days upon differentiation in N2 medium. However, in differentiation by RA/AA treatment system, there was a significant increase in proportion of neuron maturity (73%) at day 14 after N2 medium. TH#2/MB03 cells expressing TH are >90% when matured at the absence of either bDNF or TGF-$\alpha$. These results suggested that TH#2/MB03 cells could be differentiated in vitro into mature neurons by RA/AA.

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