• Title/Summary/Keyword: Cell labeling

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Development of bombesin peptide kit for prostate cancer diagnosis and treatment using 64Cu/177Lu.

  • Kim, Mi Hyun;Kim, Min Hwan;Kim, Kwang Il;Kim, Jung Young;Lee, Tae Sup;Kang, Joo Hyun;Lee, Kyo Chul;Lee, Yong Jin
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.3 no.2
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    • pp.59-64
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    • 2017
  • It has been reported that $^{64}Cu$ was radiolabeled with bombesin (BBN) peptide binding to the gastrin releasing peptide receptor expressed in human prostate cancer cells (PC3), confirming tumor target efficacy in mouse model. In this study, we developed the kit for the diagnosis and treatment of prostate cancer that can be used clinically using bombesin peptide available of $^{64}Cu$ and $^{177}Lu$ radioisotope labeling. The NODAGA-galacto-BBN peptide containing the NODAGA chelator and galactose was dispensed into a sterilized glass vial and lyophilized to prepare a kit. The stability of the kit after long-term storage in the $4^{\circ}C$ cold chamber and the radiolabeling efficiency after $^{64}Cu$ or $^{177}Lu$ labeling were confirmed by thin layer chromatography. When labeling with $^{64}Cu$ at the initial stage of storage, labeling efficiency of NODAGA-galacto-BBN peptide kit was over 96%, labeling efficiency was over 90% when $^{177}Lu$ was labeled. At 11 months after storage, the radiolabeling efficiency of kit against $^{64}Cu$ and $^{177}Lu$ was each over 95% and 90%. The cell viability was significantly reduced in the $^{177}Lu$-NODAGA-galacto-BBN treated group compared with the control and $^{177}Lu$ alone treated group in clonogenic assay. In conclusion, the NODAGA-galacto-BBN kit prepared by the lyophilization showed high stability over time and high yield of radioisotope labeling. Also $^{177}Lu$-NODAGA-galacto-BBN confirmed high cytotoxicity to prostate cancer cells. Therefore, the NODAGA-galacto-bombesin kit is expected to be useful for the diagnosis and treatment of prostate cancer patients.

The Present Status of Cell Tracking Methods in Animal Models Using Magnetic Resonance Imaging Technology

  • Kim, Daehong;Hong, Kwan Soo;Song, Jihwan
    • Molecules and Cells
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    • v.23 no.2
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    • pp.132-137
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    • 2007
  • With the advance of stem cell transplantation research, in vivo cell tracking techniques have become increasingly important in recent years. Magnetic resonance imaging (MRI) may provide a unique tool for non-invasive tracking of transplanted cells. Since the initial findings on the stem cell migration by MRI several years ago, there have been numerous studies using various animal models, notably in heart or brain disease models. In order to develop more reliable and clinically applicable methodologies, multiple aspects should be taken into consideration. In this review, we will summarize the current status and future perspectives of in vivo cell tracking technologies using MRI. In particular, use of different MR contrast agents and their detection methods using MRI will be described in much detail. In addition, various cell labeling methods to increase the sensitivity of signals will be extensively discussed. We will also review several key experiments, in which MRI techniques were utilized to detect the presence and/or migration of transplanted stem cells in various animal models. Finally, we will discuss the current problems and future directions of cell tracking methods using MRI.

Radioautographical observations of development and appearance of glia cells in brain II. Division and migration of ectodermal glial cell in the brain (뇌신경교세포(腦神經膠細胞) 집단(集團)의 발생(發生)과 이동(移動)에 대한 방사선(放射線) 자기법적(自記法的) 관찰 II. 뇌(腦) 외배엽성(外胚葉性) 신경교세포(神經膠細胞)의 분열(分裂)과 이동(移動)에 대하여)

  • Kwak, Soo-dong
    • Korean Journal of Veterinary Research
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    • v.32 no.4
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    • pp.489-496
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    • 1992
  • The division, distribution and migration of the macroglial cells in the juvenile mouse brain were investigated with the radioautography. Forty mice (ICR) were randomly subdivided into two groups. The twenty mice from group 1 were weighing initially 5 to 6g, aged 10 to 12 days and were sacrificied at 2 hrs, 2, 3, 5, 7, 10, 15 and 20 days after a single intraperitoneal injection of $^3H$-thymichine ($4{\mu}$ Ci/g of body weight). Twenty mice from group 2 were weighing intially 2.5 to 5g, aged 3 to 8 days and were sacrificed at 2 hrs, 2, 3, 5. 7, 10, 15 and 20 days after a single($4{\mu}$ Ci/g of body weight) and/or after intraperitoneal repeated injections($2{\mu}$ Ci/g of body weight/interval) at 2, 3 and 5 days after the first injection. The brain preparations were processed for autoradiogrouphy using Kodak NTB-3 emulsion following development in Kodak D-19, fixation in Kodak fixer, and then stained with cresyl echt violet or hematoxylin counterstain. The labeling index of the ectodermal glial cells in the subependymal layers of the lateral ventricles (SLLV), corpus callosum (CC), molecular layer of the neocortex (MLN ), inner layer except the molecular layer in the neocortex (ILN) and medulla of the cerebrum (MC) were invested. 1. Labeling cells appeared from 2 hour and some of them sustained in the 20 day after injection. In the single injection group, the peak of the labeling index reached a 7.6% at 3 day, 3.6% at 7 day, 3.3% at 2 day, 5.0% at 3 day and 2.3% at 2 day from the SLLV. CC, MLN, ILN and MC, respectively. In the repeated injecton group, the peak of the labeling index reached a 32.0 at 7 day, 11.0% at 10 day, 89% at 7 day, 16.0% at 10 day and 10.8% at 15 day from the SLLV, CC, MLN, ILM and MC, respectively. 2 The glial cells of the SLLV were recognized as to be migrated into the CC and to be not or less to be into the MC and ILN but to be not into the MLN. Glial cell aggregates in the neocotex and MC were recognized as to be proliferated and then disappeared in the itself regions.

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Determination of Ceramide-Induced Apoptotic Cell Death in Mouse Granulosa Cell Cultured In Vitro (생쥐 난소 과립세포의 체외배양중 세라마이드에 의한 자연세포사의 측정)

  • Kim, Jong-Hoon;Kim, Gyoung-Mi;Yoon, Yong-Dal
    • Development and Reproduction
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    • v.2 no.1
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    • pp.1-8
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    • 1998
  • In mammalian ovary, major portion(>99%) of ovarian follicles undergo atresia. Recent studies have shown that this phenomenon is mediated via GC apoptosis. Ceramide, a product of sphingomyelin hydrolysis, has been proposed as a novel lipid second messenger with specific roles in mediating antiproliferative responses including apoptosis and cell cycle arrest. In the present study, we have examined the effect of ceramide on apoptotic cell death of GC in vitro. GCs were harvested by squeezing the antral follicles from the immature mice (3-4 weeks) and cultured in MEM medium with 10% fetal bovine serum. The cells were treated with various concentrations of ceramide (0 to 50 \mu M)and cultured up to 24 h.Cell death was determined by MTT cell viability assay and apoptosis was examined by acridine orange staining, in situ 3'-end labeling(TUNEL), and flow cytometry. Ceramid treatment induced apoptotic cell death of GC in a time- and a dose-dependent manner. Results of flow cytometric analysis showed that creamide-induced cell death was mostly confined to the $G_{0}$/$G_{1}$ cells. these results provide an evidence for ceramide as a lipid second messenger of apoptosis in mouse GC.

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Expression of Kainate Glutamate Receptors in Type II Cells in Taste Buds of Rats

  • Lee, Sang-Bok;Lee, Cil-Han;Cho, Young-Kyung;Chung, Ki-Myung;Kim, Kyung-Nyun
    • International Journal of Oral Biology
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    • v.33 no.3
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    • pp.83-89
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    • 2008
  • Glutamate-induced cobalt uptake reveals non-NMDA glutamate receptors (GluRs) in rat taste bud cells. Previous studies suggest that glutamate-induced cobalt uptake in taste cells occurs mainly via kainate type GluRs. Cobaltstained cells were immunoreactive against GluR6 and KA1 subunits of GluRs. However, the functions of those type of receptors are not known yet. It is important question which types of taste cells are cobalt-stained when stimulated by glutamate and whether they express these kinds of GluRs. Circumvallate and foliate papilla of Sprague-Dawley rats (45-60 days old) were used. A cobalt-staining technique combined with immunohistochemistry against specific markers for taste bud cell types, such as blood group H antigen (BGH), $\alpha$-gustducin (Gus), or neural cell adhesion molecule (NCAM) was employed. We also performed double labeling of GluR6 or KA1 subunits of GluR with each specific marker for taste bud cell types. Lots of cobaltstained taste bud cells expressed Gus-like immunoreactivity, and subsets of the cobalt stained cells appeared NCAM- or BGH-like immunoreactivity. Stimulation with 1 mM glutamate significantly increased the number of cobaltstained cells in Gus-like immunoreactive cells, but not in NCAM- or BGH-like immunoreactive cells. In the double labeling experiments, GluR6 and KA1 subunits of GluRs were mainly expressed with Gus. These results suggest that kainate glutamate receptors preferentially expressed in type II taste bud cells in rat.

Immunoelectron Microscopic Observation on the Stratified Squamous Epithelial Cello of Pemphigus vulgaris (Pemphigus vulgaris의 중층 편평상피세포에 대한 면역전자현미경적 관찰)

  • Lee, Cha-Soo;Picut, Catherine-A.;Wilkinson, J.-Erby;Lewis, Robert-M.
    • Applied Microscopy
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    • v.17 no.2
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    • pp.1-22
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    • 1987
  • Pemphigus vulgar교의 본태(本態)를 알아보기 위한 일환으로 본질병(本疾病)의 병변조직(病變組織)을 전자현미경적(電子顯微鏡的)으로 관찰(觀察)하고 본질병(本疾病)에 관여(關與)하는 세포(細胞)들에 대(對)해 면역전자현미경적(免疫電子顯微鏡的)으로 추구(追究)하였던 바, 그 결과(結果)를 보고(報告)하는 바이다. 견(犬)의 pemphigus vulgaris의 구강(口腔) 및 식도(食道)의 점막층(粘膜層)을 전자현미경적(電子顯微鏡的)으로 관찰(觀察)하였던 바 acantholysis를 일으켜도 desmosome과 기저막(基底膜)은 큰 변화(變化)가 없었으며 세포간(細胞間)은 세포간물질(細胞間物質)의 집적(集積)에 의(依)해 확장(擴張)되고 이들 세포간강물질(細胞間腔物質)은 집괴(集塊) 또는 무구조(無構造)한 물질(物質)로 나타났다. 그리고 기저세포(基底細胞)는 기저막(基底膜)에 단단히 부착(附着)되어 있었고 dendritic cell이 기저세포층(基底細胞層)위로 분포(分布)되어 있었으며, 이들 dendritic cell 중(中)에서는 가끔 여러 형태(形態)의 퇴행성변화(退行性變化)를 볼 수 있었다. Mouse 피부유리상피세포(皮膚遊離上皮細胞)에 있어서 immunogold-labeling 방법(方法)에 의해 dendritic cell을 동정(同定)하는 데에는 post-fixation, pre-embedding immunogold-labeling technique가 좋았으며 15nm와 40nm 크기의 colloid-fold 입자(粒子)로 Langerhans cell과 Thy-1양성(陽性) dendritic cell이 표식(標識)될 수 있었다. 이들 세포(細胞)들은 세포막항원(細胞膜抗元)에 따라 monoclonal antibody의 반응(反應)에 이어 치밀한 colloid-gold 입자(粒子)가 세포막표면(細胞膜表面)을 따라 일정(一定)하게 표식(標識)되었다. 또한 이들 상피세포(上皮細胞)들을 투과전자현미경적(透過電子顯微鏡的)으로 관찰(觀察)하였을 때 초미세구조(超微細構造)가 잘 보존(保存)되었으나 Langer-hans cell내(內)의 Birbeck granule은 유리전(遊離前) 피부상피조직내(皮膚上皮組織內)의 Langerhans cell내(內)의 Birbeck granule에 비(比)해 수적(數的)으로 현저히 감소(減少)되어 있었다. 그러나 Thy-1 양성(陽性) dendritic cell에서 볼 수 있는 dense-core 과립(顆粒)은 별변화(別變化)없이 쉽게 관찰(觀察)될 수 있었다. 조직배양(組織培養)을 한 견(犬)의 keratinocyte에 대(對)해 사람 pemphigus vulgaris의 항체(抗體)로 반응(反應)시킨 후 protein-A gold(15 nm)로 표식(標識)시킨 바 제일 바깥 상층(上層)의 keratinocyte에 있어서 세포막표면(細胞膜表面)을 따라 표식(標識)되어 세포막항원(細胞膜抗元)을 나타내었으며, 이와 같은 소견(所見)으로 미루어 정상피부(正常皮膚) 중층편평상피세포(重層扁平上皮細胞)에서도 동일(同一)한 소견(所見)을 관찰(觀察)할 수 있다고 본다.

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Fucoidan Extract from Laminaria religiosa Suppresses Ischemia-induced Apoptosis and Cell Proliferation in the Hippocampus of Gerbils

  • Lee, Jong-Jin;Song, Yun-Kyung;Lim, Hyung-Ho
    • The Journal of Korean Medicine
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    • v.27 no.4
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    • pp.105-115
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    • 2006
  • Fucoidan has been shown to exhibit a host of biological activities, including anti-coagulant, anti-thrombotic, anti-tumourigenic, anti-inflammatory, anti-viral, anti-complementary and neuroprotective effects. In the present study, we attempted to determine the effects of Fucoidan on both apoptosis and cell proliferation in the hippocampal CA1 region and the dentate gyrus of gerbils after the induction of transient global ischemia. This experiment involved the use of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay as well as immunohistochemisty for caspase-3 and 5-bromo-2'-deoxyuridine (BrdU). The monosaccharide composition of the purified Fucoidan which had been extracted from Laminaria religiosa was utilized in this study. The present study clearly induces that apoptotic cell death and cell proliferation in the gerbil's hippocampal regions increased significantly following the induction of transient global ischemia and the results of this study also indicate that Fucoidan exerted a suppressive effect on this observed ischemia-induced increase in apoptosis within the CA1 and dentate gyrus, and also suppressed cell proliferation in the dentate gyrus.

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Neuroprotective Effect of Ginseng radix on ICH-induced Rats

  • Jang, Kwan-Ho;Song, Yun-Kyung;Lim, Hyung-Ho
    • The Journal of Korean Medicine
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    • v.26 no.4
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    • pp.87-97
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    • 2005
  • Backgrounds: Intracerebral hemon-hage is one of the most devastating types of stroke. Ginseng radix, the root of Panax Ginseng, C. A. MEYER (Araliaceae), is one of the most famous medicinal herbs with various therapeutic applications. Objectives: In the present study, the effect of aqueous extract of Ginseng radix on intracerebral hemorrhage-induced neuronal cell death in rats was investigated. Materials and Methods: Step-down avoidance task, Nissl staining, immunohistochemistry for caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay were used for this study. Results: The present results show that hemorrhage-induced lesion volume and apoptotic neuronal cell death in the striatum were significantly suppressed by treatment with Ginseng radix, resulting in enhancement of short-ten-n memory. Conclusions: We have shown that Ginseng radix has a neuroprotective effect on stroke, and aids the recovery from central nervous system sequelae following stroke.

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Radiolabeling and Immunological Characteristics of In-house Anti-Leukemic Monoclonal Antibodies(Anti-CALLA, Anti-JL-1 Antibodies) (국산 항 백혈병 항체(항 CALLA, 항 JL-1)의 동위원소 표지 및 면역학적 특성에 관한 연구)

  • So, Young;Chung, June-Key;Jeong, Jae-Min;Lee, Dong-Soo;Lee, Myung-Chul;Koh, Chang-Soon;Park, Seong-Hoe
    • The Korean Journal of Nuclear Medicine
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    • v.29 no.1
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    • pp.98-104
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    • 1995
  • Recently murine monoclonal antibodies have been studied actively for radioimmuno-scintigraphy and radioimmunotherapy, especially on patients with leukemia and lymphoma. In this research, we studied radiolabeling and immunologic characteristics of two in-house anti-leukemic monoclonal antibodies(anti-CALLA & anti-JL-1 antibodies) to make the basis for their clinical application. Each antibody was radiolabeled successfully with $^{99m}Tc$ by pretargeting transchelation method and with $^{125}I$ by lodogen method. We also studied cell binding assay, Scatchard analysis and modulation phenomenon. $^{125}I$ showed 90% labeling efficiency for each anti-body which was satisfactory, but $^{99m}Tc$ showed labeling efficiency below 70%, for which we need better labeling method. In cell binding assay, the immunoreactivity(IR) was low for $^{99m}Tc$-labeled antibodies. Scatchard analysis showed satisfactory data for both binding affinity. The affinity constant and antibody binding sites per cell are around $10^9M^{-1}$ and $10^4$, respectively. There was no modulation phenomenon in cases of $^{125}I$ or $^{99m}Tc$ labeled antibodies. We expect that two anti-leukemic monoclonal antibodies may be useful in diagnosis and therapy for leukemia and lymphoma patients.

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The Protective Effects of N-Acetyl-L-cysteine on Cadmium-induced Cell Apoptosis in Rat Testis

  • Kim, Ji-Sun;Soh, Jaemog
    • Biomedical Science Letters
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    • v.25 no.4
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    • pp.417-425
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    • 2019
  • Cadmium (Cd) generates reactive oxygen species (ROS), which in turn cause the apoptosis of various cell types including developing germ cells in rodent testis. Ascorbic acids (AA), one of the ROS scavengers, had been reported to protect against Cd-induced apoptosis. N-Acetyl-L-cysteine (NAC), another ROS scavenger, is known to remove ROS and alleviate the Cd-induced apoptosis in various cell types. In this study we tried to elucidate how NAC affected on Cd-induced cell apoptosis in rat testis. Rats were administered with NAC before and after Cd treatment and then testicular cell apoptosis was examined. NAC treatment resulted in the reduction of Cd-induced chromosomal DNA fragmentation in agarose gel electrophoresis. Terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) assay showed that treatment of NAC reduced the Cd-induced apoptosis of germ cells. The administration of NAC showed that the translocation of apoptosis inducing factor (AIF) from mitochondria to nucleus was prevented, which indicated that the mechanism of Cd-induced testicular apoptosis is mediated through the release of AIF in caspase-independent manner. Taken together, the NAC may remove Cd-induced ROS and protect ROS-induced cell apoptosis in rat testis.