• 제목/요약/키워드: human embryonic stem cell research

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Generation and Characterization of a Monoclonal Antibody with Specificity for Mycoplasma arginini

  • Son, Yeon-Sung;Hong, Hyo-Jeong
    • Journal of Microbiology
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    • 제45권6호
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    • pp.547-552
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    • 2007
  • Previously, we generated monoclonal antibodies (MAbs) that bound to the surface of human embryonic stem cells (hESCs) in an attempt to discover new hESC-specific surface markers. In this study, MAb 47-235 (IgG1, ${\kappa}$) was selected for further characterization. The MAb bound to the surface of undifferentiated hESCs but did not bind to mouse ESCs or mouse embryonic fibroblast cells in flow cytometric analysis. The antibody immunoprecipitated a 47 kDa protein from the lysates of cell surface-biotinylated hESCs. Identification of the protein by quadrupole time of flight tandem mass spectrometry revealed that 47-235 binds to Ag 243-5 protein of Mycoplasma arginini. BM-Cyclin treatment of the hESCs that reacted with 47-235 resulted in loss of mycoplasma DNA and the reactivity to 47-235. Nevertheless, the hESCs that were reactive to 47-235 maintained self-renewal and pluripotency and thus could be differentiated into three embryonic germ layers.

Expression of HERV-HX2 in Cancer Cells and Human Embryonic Stem Cells

  • Jung, Hyun-Min;Choi, Seoung-Jun;Kim, Se-Hee;Moon, Sung-Hwan;Yoo, Jung-Ki;Chung, Hyung-Min;Kim, Jin-Kyeoung
    • Reproductive and Developmental Biology
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    • 제32권2호
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    • pp.105-110
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    • 2008
  • The endogenous retrovirus-like elements (HERVs) found on several human chromosomes are somehow involved in gene regulation, especially during the transcription level. HERV-H, located on chromosome Xp22, may regulate gastrin-releasing peptide receptor (GRPR) in connection with diverse diseases. By suppression subtractive hybridization screen on SV40-immortalized lung fibroblast (WI-38 VA-13), we discovered that expression of HERV-HX2, a clustered HERV-H sequence on chromosome X, was upregulated in immortalized lung cells, compared to that of normal cells. Expression of HERV-HX2 was then analyzed in various cell lines, including normal somatic cells, cancer cells, SV40-immortalized cells, and undifferentiated and differentiated human embryonic stem cells. Expression of HERV-HX2 was specifically upregulated in continuously-dividing cells, such as cancer cells and SV40-immortalized cells. Especially, HERV-HX2 in HeLa cells was highly upregulated during the S phase of the cell cycle. Similar results were obtained in hES cells, in which undifferentiated cells expressed more HERV-HX2 mRNA than differentiated hES cells, including neural precursor and endothelial progenitor cells. Taken together, our results suggest that HERV-HX2 is upregulated in cancer cells and undifferentiated hES cells, whereas downregulated as differentiation progress. Therefore, we assume that HERV-HX2 may playa role on proliferation of cancer cells as well as differentiation of hES cells in the transcriptional level.

동결에 앞서 시행된 지지세포와의 추가 공배양이 인간 배아줄기세포의 유리화 동결보존 후 생존율과 미분화 유지에 미치는 영향 (Improving the Survival and Maintenance of the Undifferentiated State of Cryopreserved Human Embryonic Stem Cells by Extended Incubation with Feeder Cells Overnight before Vitrification)

  • 차수경;최경희;신주미;박규형;윤태기;정형민;이동률
    • 한국발생생물학회지:발생과생식
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    • 제12권2호
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    • pp.141-149
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    • 2008
  • This study was conducted to develop an efficient cryopreservation method of human embryonic stem (ES) cells using vitrification. In an initial experiment, sub-clumps of human ES cells (CHA-hES3 and CHA-hES4) were vitrified using grids after incubation with STO feeder cells for 1 or 16 h (Groups 1-1 and 1-2, respectively). After storage for $2{\sim}4$ months, thawed clumps were re-plated on a fresh feeder layer. The survival rates of warmed CHA-hES3 and CHA-hES4 cells of Group 1-2 were significantly higher than those of the corresponding Group 1-1 cells. In the second experiment, human ES cells were vitrified after incubation with feeder or feeder-conditioned medium (Groups 2-1 to -7). Relative mRNA expression of BM proteins and survival rates were increased following incubation of ES cells with fresh feeder cells for 16 h. In conclusion, increasing of tight adhesion between ES cells by extended incubation with feeder could reduce cryoinjury after vitrifying/warming.

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Assessment of Developmental Toxicants using Human Embryonic Stem Cells

  • Hong, Eui-Ju;Jeung, Eui-Bae
    • Toxicological Research
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    • 제29권4호
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    • pp.221-227
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    • 2013
  • Embryonic stem (ES) cells have potential for use in evaluation of developmental toxicity because they are generated in large numbers and differentiate into three germ layers following formation of embryoid bodies (EBs). In earlier study, embryonic stem cell test (EST) was established for assessment of the embryotoxic potential of compounds. Using EBs indicating the onset of differentiation of mouse ES cells, many toxicologists have refined the developmental toxicity of a variety of compounds. However, due to some limitation of the EST method resulting from species-specific differences between humans and mouse, it is an incomplete approach. In this regard, we examined the effects of several developmental toxic chemicals on formation of EBs using human ES cells. Although human ES cells are fastidious in culture and differentiation, we concluded that the relevancy of our experimental method is more accurate than that of EST using mouse ES cells. These types of studies could extend our understanding of how human ES cells could be used for monitoring developmental toxicity and its relevance in relation to its differentiation progress. In addition, this concept will be used as a model system for screening for developmental toxicity of various chemicals. This article might update new information about the usage of embryonic stem cells in the context of their possible ability in the toxicological fields.

In vitro neural differentiation of human embryonic stem cells

  • Park, Jae-Hyun;Shin, Hwa-Yean;Kang, Yun-Hee;Kang, Young-Kook;Lee, Jung-Bok;Yoon, Hyun-Soo;Ryu, Chun-Jeih;Myung, Pyung-Keun;Hong, Hyo-Jeong
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.164.2-164.2
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    • 2003
  • Human embryonic stem (ES) cell lines derived from the inner cell mass of human blastocysts have potential to differentiate into any cell types. We have established in vitro neural differentiation of human ES cells. After the formation of embroid bodies (EBs), the differentiating EBs formed neural tube-like rosettes in the presence of basic fibroblast growth factor (bFGF). The rosettes were selectively isolated by the treatment of dispase and cultured in a medium for human neural precursors in the presence of bFGF. (omitted)

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Epigenetic modification of retinoic acid-treated human embryonic stem cells

  • Cheong, Hyun-Sub;Lee, Han-Chul;Park, Byung-Lae;Kim, Hye-Min;Jang, Mi-Jin;Han, Yong-Mahn;Kim, Seun-Young;Kim, Yong-Sung;Shin, Hyoung-Doo
    • BMB Reports
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    • 제43권12호
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    • pp.830-835
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    • 2010
  • Epigenetic modification of the genome through DNA methylation is the key to maintaining the differentiated state of human embryonic stem cells (hESCs), and it must be reset during differentiation by retinoic acid (RA) treatment. A genome-wide methylation/gene expression assay was performed in order to identify epigenetic modifications of RA-treated hESCs. Between undifferentiated and RA-treated hESCs, 166 differentially methylated CpG sites and 2,013 differentially expressed genes were discovered. Combined analysis of methylation and expression data revealed that 19 genes (STAP2, VAMP8, C10orf26, WFIKKN1, ELF3, C1QTNF6, C10orf10, MRGPRF, ARSE, LSAMP, CENTD3, LDB2, POU5F1, GSPT2, THY1, ZNF574, MSX1, SCMH1, and RARB) were highly correlated with each other. The results provided in this study will facilitate future investigations into the interplay between DNA methylation and gene expression through further functional and biological studies.

Cancer stem cell surface markers on normal stem cells

  • Kim, Won-Tae;Ryu, Chun Jeih
    • BMB Reports
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    • 제50권6호
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    • pp.285-298
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    • 2017
  • The cancer stem cell (CSC) hypothesis has captured the attention of many scientists. It is believed that elimination of CSCs could possibly eradicate the whole cancer. CSC surface markers provide molecular targeted therapies for various cancers, using therapeutic antibodies specific for the CSC surface markers. Various CSC surface markers have been identified and published. Interestingly, most of the markers used to identify CSCs are derived from surface markers present on human embryonic stem cells (hESCs) or adult stem cells. In this review, we classify the currently known 40 CSC surface markers into 3 different categories, in terms of their expression in hESCs, adult stem cells, and normal tissue cells. Approximately 73% of current CSC surface markers appear to be present on embryonic or adult stem cells, and they are rarely expressed on normal tissue cells. The remaining CSC surface markers are considerably expressed even in normal tissue cells, and some of them have been extensively validated as CSC surface markers by various research groups. We discuss the significance of the categorized CSC surface markers, and provide insight into why surface markers on hESCs are an attractive source to find novel surface markers on CSCs.

Porcine OCT4 reporter system as a tool for monitoring pluripotency states

  • Kim, Seung-Hun;Lee, Chang-Kyu
    • 한국동물생명공학회지
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    • 제36권4호
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    • pp.175-182
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    • 2021
  • Pluripotent stem cells could self-renew and differentiate into various cells. In particular, porcine pluripotent stem cells are useful for preclinical therapy, transgenic animals, and agricultural usage. These stem cells have naïve and primed pluripotent states. Naïve pluripotent stem cells represented by mouse embryonic stem cells form chimeras after blastocyst injection. Primed pluripotent stem cells represented by mouse epiblast stem cells and human embryonic stem cells. They could not produce chimeras after blastocyst injection. Populations of embryonic stem cells are not homogenous; therefore, reporter systems are used to clarify the status of stem cells and to isolate the cells. For this reason, studies of the OCT4 reporter system have been conducted for decades. This review will discuss the naïve and primed pluripotent states and recent progress in the development of porcine OCT4 reporter systems.