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Characterization of common marmoset dysgerminoma-like tumor induced by the lentiviral expression of reprogramming factors.

Yamaguchi S, Marumoto T, Nii T, Kawano H, Liao J, Nagai Y, Okada M, Takahashi A, Inoue H, Sasaki E, Fujii H, Okano S, Ebise H, Sato T, Suyama M, Okano H, Miura Y, Tani K - Cancer Sci. (2014)

Bottom Line: Recent generation of induced pluripotent stem (iPSCs) has made a significant impact on the field of human regenerative medicine.Prior to the clinical application of iPSCs, testing of their safety and usefulness must be carried out using reliable animal models of various diseases.In order to generate iPSCs from common marmoset (CM; Callithrix jacchus), one of the most useful experimental animals, we have lentivirally transduced reprogramming factors, including POU5F1 (also known as OCT3/4), SOX2, KLF4, and c-MYC into CM fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular and Clinical Genetics, Molecular and Clinical Genetics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.

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Chromosome abnormality and tumor-forming ability in abnormally reprogrammed cells (ARCs). (a) Karyotype analyses of aorta-gonado-mesonephros (AGM) fibroblasts (left panel) and ARCs (right panel). Arrows indicate marker chromosome. Blue outline indicates the deletion of 4q. Mar, marker chromosome. (b) Representative photograph of dysgerminoma-like tumor (arrow) formed by transplantation of ARCs into SCID mice. (c) Hematoxylin-eosin staining of dysgerminoma-like tumor tissues. Arrows in right panel indicate mitotic figures in tumor cells. Bar = 100 μm. (d) Microarray analysis. Gene expressions in AGM fibroblasts, ARCs, and normal induced pluripotent stem (iPS) A cells were analyzed by unsupervised hierarchical clustering. A heat map using probes showing differential expression levels in each cell line is shown. Red indicates upregulation; green indicates downregulation. The black bar on the right side of the heat map shows candidate differentially expressed probes in ARCs.
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fig02: Chromosome abnormality and tumor-forming ability in abnormally reprogrammed cells (ARCs). (a) Karyotype analyses of aorta-gonado-mesonephros (AGM) fibroblasts (left panel) and ARCs (right panel). Arrows indicate marker chromosome. Blue outline indicates the deletion of 4q. Mar, marker chromosome. (b) Representative photograph of dysgerminoma-like tumor (arrow) formed by transplantation of ARCs into SCID mice. (c) Hematoxylin-eosin staining of dysgerminoma-like tumor tissues. Arrows in right panel indicate mitotic figures in tumor cells. Bar = 100 μm. (d) Microarray analysis. Gene expressions in AGM fibroblasts, ARCs, and normal induced pluripotent stem (iPS) A cells were analyzed by unsupervised hierarchical clustering. A heat map using probes showing differential expression levels in each cell line is shown. Red indicates upregulation; green indicates downregulation. The black bar on the right side of the heat map shows candidate differentially expressed probes in ARCs.

Mentions: Total RNA from AGM fibroblasts, ARCs, and iPS A cells were isolated using the RNeasy Mini Kit (Qiagen, Valencia, CA, USA). RNA was reverse-transcribed, biotin-labeled, and hybridized for 16 h to a marmoset genome oligonucleotide custom array Marmo2 (in preparation)(12), which was subsequently washed and stained in a Fluidics Station 450 (Affymetrix, Santa Clara, CA, USA) according to the manufacturer's instructions. Detailed protocols of microarray analysis are provided in Figures 2 and S5.


Characterization of common marmoset dysgerminoma-like tumor induced by the lentiviral expression of reprogramming factors.

Yamaguchi S, Marumoto T, Nii T, Kawano H, Liao J, Nagai Y, Okada M, Takahashi A, Inoue H, Sasaki E, Fujii H, Okano S, Ebise H, Sato T, Suyama M, Okano H, Miura Y, Tani K - Cancer Sci. (2014)

Chromosome abnormality and tumor-forming ability in abnormally reprogrammed cells (ARCs). (a) Karyotype analyses of aorta-gonado-mesonephros (AGM) fibroblasts (left panel) and ARCs (right panel). Arrows indicate marker chromosome. Blue outline indicates the deletion of 4q. Mar, marker chromosome. (b) Representative photograph of dysgerminoma-like tumor (arrow) formed by transplantation of ARCs into SCID mice. (c) Hematoxylin-eosin staining of dysgerminoma-like tumor tissues. Arrows in right panel indicate mitotic figures in tumor cells. Bar = 100 μm. (d) Microarray analysis. Gene expressions in AGM fibroblasts, ARCs, and normal induced pluripotent stem (iPS) A cells were analyzed by unsupervised hierarchical clustering. A heat map using probes showing differential expression levels in each cell line is shown. Red indicates upregulation; green indicates downregulation. The black bar on the right side of the heat map shows candidate differentially expressed probes in ARCs.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4317795&req=5

fig02: Chromosome abnormality and tumor-forming ability in abnormally reprogrammed cells (ARCs). (a) Karyotype analyses of aorta-gonado-mesonephros (AGM) fibroblasts (left panel) and ARCs (right panel). Arrows indicate marker chromosome. Blue outline indicates the deletion of 4q. Mar, marker chromosome. (b) Representative photograph of dysgerminoma-like tumor (arrow) formed by transplantation of ARCs into SCID mice. (c) Hematoxylin-eosin staining of dysgerminoma-like tumor tissues. Arrows in right panel indicate mitotic figures in tumor cells. Bar = 100 μm. (d) Microarray analysis. Gene expressions in AGM fibroblasts, ARCs, and normal induced pluripotent stem (iPS) A cells were analyzed by unsupervised hierarchical clustering. A heat map using probes showing differential expression levels in each cell line is shown. Red indicates upregulation; green indicates downregulation. The black bar on the right side of the heat map shows candidate differentially expressed probes in ARCs.
Mentions: Total RNA from AGM fibroblasts, ARCs, and iPS A cells were isolated using the RNeasy Mini Kit (Qiagen, Valencia, CA, USA). RNA was reverse-transcribed, biotin-labeled, and hybridized for 16 h to a marmoset genome oligonucleotide custom array Marmo2 (in preparation)(12), which was subsequently washed and stained in a Fluidics Station 450 (Affymetrix, Santa Clara, CA, USA) according to the manufacturer's instructions. Detailed protocols of microarray analysis are provided in Figures 2 and S5.

Bottom Line: Recent generation of induced pluripotent stem (iPSCs) has made a significant impact on the field of human regenerative medicine.Prior to the clinical application of iPSCs, testing of their safety and usefulness must be carried out using reliable animal models of various diseases.In order to generate iPSCs from common marmoset (CM; Callithrix jacchus), one of the most useful experimental animals, we have lentivirally transduced reprogramming factors, including POU5F1 (also known as OCT3/4), SOX2, KLF4, and c-MYC into CM fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular and Clinical Genetics, Molecular and Clinical Genetics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.

Show MeSH
Related in: MedlinePlus