• Title/Summary/Keyword: Cell lineage

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A Study on the Hierarchical Expression of Human Cell Lineage (인간 세포 Lineage 의 계층적 표현에 관한 연구)

  • Park, JaeSoon;Kwon, Seong Gyu;Oh, Ji Won;Lee, JongHyuk
    • Proceedings of the Korea Information Processing Society Conference
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    • 2020.11a
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    • pp.663-664
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    • 2020
  • 차세대 염기서열 분석 기술은 성능과 비용 면에서 매우 향상되어 한 개체 내 여러 세포의 유전자 분석이 가능한 수준이다. 한 개체 내 여러 조직 세포의 유전자는 모두 동일하지 않기 때문에 여러 조직 세포의 Lineage 를 계층적으로 표현하고 이를 조직 세포 간 변이 정도를 파악하는 데 활용한다면 암 돌연변이 발생 등을 미리 예측할 수 있다. 본 논문은 한 개체 내 여러 조직 간 변이를 관찰하기 위해 변이 검출 데이터를 계층적 군집 방법을 이용해 분석하고 이를 시각화 하는 방법을 제안한다. 실제의 8 개 조직 세포의 유전자를 분석하고 변이를 검출하여 Dendrogram 그래프로 시각화 하였다.

Comparative Analysis for In Vitro Differentiation Potential of Induced Pluripotent Stem Cells, Embryonic Stem Cells, and Multipotent Spermatogonial Stem Cells into Germ-lineage Cells

  • Go, Young-Eun;Kim, Hyung-Joon;Jo, Jung-Hyun;Lee, Hyun-Ju;Do, Jeong-Tae;Ko, Jung-Jae;Lee, Dong-Ryul
    • Development and Reproduction
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    • v.15 no.1
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    • pp.41-52
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    • 2011
  • In the present study, embryoid bodies (EBs) obtained from induced pluripotent stem cells (iPSCs) were induced to differentiate into germ lineage cells by treatment with bone morphogenetic protein 4 (BMP4) and retinoic acid (RA). The results were compared to the results for embryonic stem cells (ESCs) and multipotent spermatogonial stem cells (mSSCs) and quantified using immunocytochemical analysis of germ cell-specific markers (integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1), fluorescence activating cell sorting (FACS), and real time-RT-PCR. We show that the highest levels of germ cell marker-expressing cells were obtained from groups treated with 10 ng/$m{\ell}$ BMP4 or 0.01 ${\mu}M$ RA. In the BMP4-treated group, GFR-${\alpha}1$ and CD90/Thy-1 were highly expressed in the EBs of iPSCs and ESCs compared to EBs of mSSCs. The expression of Nanog was much lower in iPSCs compared to ESCs and mSSCs. In the RA treated group, the level of GFR-${\alpha}1$ and CD90/Thy-1 expression in the EBs of mSSCs Induced pluripotent stem cells, Mouse embryonic stem cells, Multipotent spermatogonial stem cells, Germ cell lineage, Differentiation potential. was much higher than the levels found in the EBs of iPSCs and similar to the levels found in the EBs of ESCs. FACS analysis using integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1 and immunocytochemistry using GFR-${\alpha}1$ antibody showed similar gene expression results. Therefore our results show that iPSC has the potential to differentiate into germ cells and suggest that a protocol optimizing germ cell induction from iPSC should be developed because of their potential usefulness in clinical applications requiring patient-specific cells.

The Rat Myosin Light Chain Promoter-Driven DsRed Reporter System Allows Specific Monitoring of Bone Marrow Mesenchymal Stem Cell- Derived Cardiomyocytes

  • Choi, Seung-Cheol;Lim, Do-Sun
    • Reproductive and Developmental Biology
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    • v.32 no.1
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    • pp.21-25
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    • 2008
  • Bone marrow mesenchymal stem cells (BMMSCs) have the capacity for self-renewal and differentiation into a variety of cell types. They represent an attractive source of cells for gene and cell therapy. The purpose of this study is to direct the specific expression of the DsRed reporter gene in $Sca-1^+$ BMMSCs differentiated into a cardiomyogenic lineage. We constructed the prMLC-2v-DsRed vector expressing DsRed under the control of the 309 tp fragment of the rat MLC-2v 5'-flanking region. The specific expression of the DsRed reporter gene under the transcriptional control of the 309 bp fragment of the rat MLC-2v promoter was tested in 5-azacytidine healed-$Sca-1^+$ BMMSCs over 2 weeks after the prMLC-2v-DsRed transfection. The prMLC-2v-DsRed was specifically expressed in the $Sca-1^+$ BMMSCs with cardiomyogenic lineage differentiation and it demonstrates that the 309 bp sequences of the rat MLC-2v 5'-flanking region is sufficient to confer cardiac specific expression on a DsRed reporter gene. The cardiac-specific promoter-driven reporter vector provides an important tool for the study of stem cell differentiation and cell replacement therapy in ischemic cardiomyopathy.

Hepatocyte Growth Factor is the Key Cytokine in Stimulating Potential Stem Cells in the Cord Blood into Hepatic Lineage Cells

  • Ryu, Kyung-Ha;Cho, Su-Jin;Woo, So-Youn;Seoh, Ju-Young;Jung, Yun-Jae;Han, Ho-Seong
    • IMMUNE NETWORK
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    • v.7 no.3
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    • pp.117-123
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    • 2007
  • Background: This study was designed to investigate the role of the hepatocyte growth factor (HGF) with regards to differentiation of somatic stem cells originating from the human umbilical cord blood (UCB) into hepatic lineage cells in vitro culture system. Methods: Mononuclear cells from UCB were cultured with and without HGF based on the fibroblast growth factor (FGF)-1, FGF-2, and stem cell factor. The cultured cells were confirmed by immunofluorescent staining analysis with albumin (ALB), cytokeratin-19 (CK-19), and proliferating cell nuclear antigen (PCNA) MoAb. ALB and CK-18 mRNA were also evaluated by reverse transcription-polymerase chain reaction. In order to observe changes in proliferating capacity with respect to the cultured period, CFSE with affinity to proliferating cells were tagged and later underwent flow cytometry. Results: In the HGF-treated group, cultured cells had a large oval shaped appearance with adherent, but easily detachable characteristics. In the HGF-non treated group, these cells were spindle-shaped with strong adherent characteristics. Expressions of ALB and CK-19 were evident in HGF-treated group compared to non-expression of those in to HGF-non treated group. Dual immunostaining analysis of the ALB producing cells showed presence of PCNA in their nuclei, and ALB and CK-18 mRNA were detected on the 21st day of cultured cells in the HGF-treated group. Conclusion: Our findings suggest that HGF has a pivotal role in differentiating somatic stem cells of human UCB into hepatic lineage cells in vitro.

Panax Ginseng Rg1 Enhances CD4+ T Cell Activities and Modulates Th1/Th2 Differentiation (인삼 Saponin Rg1이 분화된 보조 T cell의 cytokine 분비에 미치는 영향)

  • Kwon Hong Rho;Ko Eun Jung;Bae Hyun Su;Hong Moo Chang;Jung Seung Gi;Shin Min Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.4
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    • pp.1021-1027
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    • 2004
  • Panax ginseng has been used as a typical tonic medicine in Asian countries, such as Korea, China, and Japan. It has been reported that ginsenoside Rg1 in Panax ginseng increases the proportion of T helper cells in the whole T cells and promotes IL-2 gene expression in murine splenocytes. These studies imply that ginsenoside Rg1 increases the immune activity of CD4+ T cell, however the exact mechanism of ginsenoside Rg1 on helper T cell remains to be verified. The present study tried to elucidate the direct effect of Rg1 on helper T cell s activities and its Th1/Th2 lineage development. The results demonstrated that ginsenoside Rg1 had not mitogenic effects on the unstimulated CD4+ T cell, but augmented CD4+ T cell proliferation upon activating with anti-CD3/anti-CD28 antibodies in a dose dependent manner. Rg1 also enhanced the expression of cell surface protein CD69 on CD4+ T cell. In Th0 condition, ginsenoside Rg1 increases the expression of IL-2 mRNA, and enhances the expression of IL-4 mRNA on CD4+ T cells, suggesting Rg1 prefer to induce Th2 lineage development. In addition, ginsenoside Rg1 increases IL-4 secreting CD4+ T cell under Th2 skewed condition, while decreases IFN-γ secreting cell in Th1 polarizing condition. Thus, Rg1 enhances Th2 lineage development from naive CD4+ T cell both by increasing Th2 specific cytokine secretion and by repressing Th1 specific cytokine production. Therefore, these results suggest that ginsenoside Rg1 might be desirable agent for enhancing CD4+ T cell's activity, as well as the correction of Th1 dominant pathological disorders.

Screening and Isolation of the Cell Differentiation Inducers from Medicinal Plants (I) (생약으로부터 세포분화유도물질의 검색 및 분리 및 분리 (I))

  • Park, Eun-Jung;Kim, Jin-Woong
    • Korean Journal of Pharmacognosy
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    • v.28 no.4
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    • pp.225-232
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    • 1997
  • 300 extracts derived from 100 plants were tested for their potential to induce HL-60 cell differentiation using NBT assay and NSE/SE staining methods. Morphological changes from suspended to adherent state of the cells were also observed by microscopic examination. In result, 55 extracts induced cell differentiation into monocyte/macrophage lineage in the NBT and the NSE assay.

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Determination of Cell Fate for Inner Cell Mass and Trophectoderm Cells in Bovine Early Cleaving Embryos (소 수정란의 초기 난할 시기에 내부 세포괴와 영양막 세포로의 운명 결정)

  • Song, Bong-Seok;Kim, Ji-Su;Kim, Cheol-Hee;Lee, Kyung-Kwang;Koo, Deog-Bon
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.193-198
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    • 2007
  • The present study was examined the expression patterns of cdx2 gone, n lineage marker, in the mouse and bovine developmental stage embryos and whether one blastomere of two- and/or four-cell bovine embryos develop to specific lineage (ICM or TE) of blastocyst by injection of Texas red conjugated dextran as a lineage tracer. It was also investigated the allocation of ICM and n cells in bovine blastocysts derived from one blastomere of two-and/or four-cell stage embryos. Firstly, it was observed that expression of cdx2 appeared symmetric and asymmetric distribution at the two-cell stage mouse embryos. from four-cell to morula stage mouse embryos, the expression of cdx2 gene was observed in almost all blastomeres. In case of bovine embryos, localization of cdx2 was similar to pattern of mouse embryos. The Dextran-labeled blastomere of two- and/or four-cell embryos contributed to both ICM and TE cells in bovine blastocysts. And also, it was confirmed that a single blastomere derived from two-cell stage bovine embryos could develop to the normal blastocyst with both ICM and TE cells. These results show that two-and/or four-cell stage is not the specific stage to determine the cell rate for ICM and TE, and which is not correlated with the expression of cdx2 gene.

The increased expression of CD99 in a differentiated neuroblastoma cell line (신경모세종의 분화에 따른 인체 CD99의 표현의 증가)

  • Choi, Eun Young;Lee, Im-Soon
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.53-60
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    • 2001
  • Background: The human mic2 gene is a pseudoautosomal gene that encodes a cell surface antigen, CD99. High levels of CD99 constitute a tumor marker in Ewing s sarcoma (ES). We have recently demonstrated that CD99-induced apoptosis occurs only in undifferentiated ES cells, not in differentiated ES cells, raising the possibility of the involvement of CD99 in neural ontogeny. Methods: To elucidate the relations between the expression of CD99 and the differentiation of neural cells and the mechanism by which the expression of CD99 is regulated, we analyzed the differential patterns of CD99 expression in SH-SY5Y by treatment of 12-O-tetradecanoyl-13-phorbol acetate (TPA) and retinoic acid. In addition, to explore the transcriptional activity of CD 99 during neural cell differentiation, SH-SY5Y cells were transiently transfected with a CD99 promoter-driven luciferase construct, and treated with the inducers. Results: In immunoblotting and flow cytometry, the expression level of CD99 was increased on differentiated SH-SY5Y cells induced by TPA and retinoic acid. The luciferase activity was elevated by the treatment with TPA, known to mature SH-SY5Y cells toward a sympathetic neuronal lineage, whereas retinoic acid inducing a sympathetic chromaffin lineage displayed little effect. Conclusion: The result indicates that CD99 might be expressed only on cells maturing toward a neuronal lineage among differentiating primitive neuronal cells. In addition, the expression of CD99 seems to be regulated at the transcriptional level during the differentiation.

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