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New type of Sendai virus vector provides transgene-free iPS cells derived from chimpanzee blood.

Fujie Y, Fusaki N, Katayama T, Hamasaki M, Soejima Y, Soga M, Ban H, Hasegawa M, Yamashita S, Kimura S, Suzuki S, Matsuzawa T, Akari H, Era T - PLoS ONE (2014)

Bottom Line: Using TS12KOS, we established iPSC lines from chimpanzee blood, and used DNA array analysis to show that the global gene-expression pattern of chimpanzee iPSCs is similar to those of human embryonic stem cell and iPSC lines.These results demonstrated that our new vector is useful for generating iPSCs from the blood cells of both human and chimpanzee.In addition, the chimpanzee iPSCs are expected to facilitate unique studies into human physiology and disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.

ABSTRACT
Induced pluripotent stem cells (iPSCs) are potentially valuable cell sources for disease models and future therapeutic applications; however, inefficient generation and the presence of integrated transgenes remain as problems limiting their current use. Here, we developed a new Sendai virus vector, TS12KOS, which has improved efficiency, does not integrate into the cellular DNA, and can be easily eliminated. TS12KOS carries KLF4, OCT3/4, and SOX2 in a single vector and can easily generate iPSCs from human blood cells. Using TS12KOS, we established iPSC lines from chimpanzee blood, and used DNA array analysis to show that the global gene-expression pattern of chimpanzee iPSCs is similar to those of human embryonic stem cell and iPSC lines. These results demonstrated that our new vector is useful for generating iPSCs from the blood cells of both human and chimpanzee. In addition, the chimpanzee iPSCs are expected to facilitate unique studies into human physiology and disease.

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Characterization of chimpanzee iPSCs.(a) Differentiation into three germ layers in vitro. The chimpanzee iPSC lines can generate SOX17+ (endoderm), BRACHYURY+ (mesoderm), and βIII-tubulin+ (ectoderm) cells. Scale bars, 100 µm. (b) Tissue morphology of a representative teratoma derived from the chimpanzee iPSC lines generated with TS12KOS vector. G, glandular structure (endoderm); C, cartilage (mesoderm); CE, Cuboidal Epithelium structure (ectoderm); MP, melanin pigment (ectoderm). Scale bars, 100 µm. (c) Principal Component Analysis. All data sets were classified into three principal components, PC1 (47.62%), PC2 (29.81%), and PC3 (22.56%), and then simplified into three-dimensional scores. Percentage shows the portion of variance in each component. The position of chimpanzee iPSC lines is closely placed to that of human ESCs and iPSCs. (d) Hierarchical clustering of chimpanzee iPSCs, human iPSCs and ESCs. The data sets of all genes investigated were clustered according to Euclidean distance metrics. The data sets of chimpanzee iPSCs, human ESC and iPSC lines, and various human tissues were classified into separate branches. The datasets of human ESCs and various tissues referred for Gene Expression Omnibus datasets, GSE22167 and GSE33846, respectively.
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pone-0113052-g004: Characterization of chimpanzee iPSCs.(a) Differentiation into three germ layers in vitro. The chimpanzee iPSC lines can generate SOX17+ (endoderm), BRACHYURY+ (mesoderm), and βIII-tubulin+ (ectoderm) cells. Scale bars, 100 µm. (b) Tissue morphology of a representative teratoma derived from the chimpanzee iPSC lines generated with TS12KOS vector. G, glandular structure (endoderm); C, cartilage (mesoderm); CE, Cuboidal Epithelium structure (ectoderm); MP, melanin pigment (ectoderm). Scale bars, 100 µm. (c) Principal Component Analysis. All data sets were classified into three principal components, PC1 (47.62%), PC2 (29.81%), and PC3 (22.56%), and then simplified into three-dimensional scores. Percentage shows the portion of variance in each component. The position of chimpanzee iPSC lines is closely placed to that of human ESCs and iPSCs. (d) Hierarchical clustering of chimpanzee iPSCs, human iPSCs and ESCs. The data sets of all genes investigated were clustered according to Euclidean distance metrics. The data sets of chimpanzee iPSCs, human ESC and iPSC lines, and various human tissues were classified into separate branches. The datasets of human ESCs and various tissues referred for Gene Expression Omnibus datasets, GSE22167 and GSE33846, respectively.

Mentions: Finally, we investigated the differentiation potential of the chimpanzee iPSCs lines by evaluating in vitro differentiation and teratoma formation (Fig. 4a, b). The appropriate culture conditions induced differentiation into cells representing all three germ layers, ectoderm (βIII-tubulin-positive), mesoderm (BRACHYURY-positive), and endoderm (SOX17-positive) (Fig. 4a). Consistent with this finding, histological analysis revealed that formed teratomas contained descendant markers of all three germ cell layers such as cuboidal epithelia, melanin pigment, cartilage, muscle, and various glandular structures (Fig. 4b). Taken together, the chimpanzee-derived iPSC lines fulfilled the criteria for iPSCs.


New type of Sendai virus vector provides transgene-free iPS cells derived from chimpanzee blood.

Fujie Y, Fusaki N, Katayama T, Hamasaki M, Soejima Y, Soga M, Ban H, Hasegawa M, Yamashita S, Kimura S, Suzuki S, Matsuzawa T, Akari H, Era T - PLoS ONE (2014)

Characterization of chimpanzee iPSCs.(a) Differentiation into three germ layers in vitro. The chimpanzee iPSC lines can generate SOX17+ (endoderm), BRACHYURY+ (mesoderm), and βIII-tubulin+ (ectoderm) cells. Scale bars, 100 µm. (b) Tissue morphology of a representative teratoma derived from the chimpanzee iPSC lines generated with TS12KOS vector. G, glandular structure (endoderm); C, cartilage (mesoderm); CE, Cuboidal Epithelium structure (ectoderm); MP, melanin pigment (ectoderm). Scale bars, 100 µm. (c) Principal Component Analysis. All data sets were classified into three principal components, PC1 (47.62%), PC2 (29.81%), and PC3 (22.56%), and then simplified into three-dimensional scores. Percentage shows the portion of variance in each component. The position of chimpanzee iPSC lines is closely placed to that of human ESCs and iPSCs. (d) Hierarchical clustering of chimpanzee iPSCs, human iPSCs and ESCs. The data sets of all genes investigated were clustered according to Euclidean distance metrics. The data sets of chimpanzee iPSCs, human ESC and iPSC lines, and various human tissues were classified into separate branches. The datasets of human ESCs and various tissues referred for Gene Expression Omnibus datasets, GSE22167 and GSE33846, respectively.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4257541&req=5

pone-0113052-g004: Characterization of chimpanzee iPSCs.(a) Differentiation into three germ layers in vitro. The chimpanzee iPSC lines can generate SOX17+ (endoderm), BRACHYURY+ (mesoderm), and βIII-tubulin+ (ectoderm) cells. Scale bars, 100 µm. (b) Tissue morphology of a representative teratoma derived from the chimpanzee iPSC lines generated with TS12KOS vector. G, glandular structure (endoderm); C, cartilage (mesoderm); CE, Cuboidal Epithelium structure (ectoderm); MP, melanin pigment (ectoderm). Scale bars, 100 µm. (c) Principal Component Analysis. All data sets were classified into three principal components, PC1 (47.62%), PC2 (29.81%), and PC3 (22.56%), and then simplified into three-dimensional scores. Percentage shows the portion of variance in each component. The position of chimpanzee iPSC lines is closely placed to that of human ESCs and iPSCs. (d) Hierarchical clustering of chimpanzee iPSCs, human iPSCs and ESCs. The data sets of all genes investigated were clustered according to Euclidean distance metrics. The data sets of chimpanzee iPSCs, human ESC and iPSC lines, and various human tissues were classified into separate branches. The datasets of human ESCs and various tissues referred for Gene Expression Omnibus datasets, GSE22167 and GSE33846, respectively.
Mentions: Finally, we investigated the differentiation potential of the chimpanzee iPSCs lines by evaluating in vitro differentiation and teratoma formation (Fig. 4a, b). The appropriate culture conditions induced differentiation into cells representing all three germ layers, ectoderm (βIII-tubulin-positive), mesoderm (BRACHYURY-positive), and endoderm (SOX17-positive) (Fig. 4a). Consistent with this finding, histological analysis revealed that formed teratomas contained descendant markers of all three germ cell layers such as cuboidal epithelia, melanin pigment, cartilage, muscle, and various glandular structures (Fig. 4b). Taken together, the chimpanzee-derived iPSC lines fulfilled the criteria for iPSCs.

Bottom Line: Using TS12KOS, we established iPSC lines from chimpanzee blood, and used DNA array analysis to show that the global gene-expression pattern of chimpanzee iPSCs is similar to those of human embryonic stem cell and iPSC lines.These results demonstrated that our new vector is useful for generating iPSCs from the blood cells of both human and chimpanzee.In addition, the chimpanzee iPSCs are expected to facilitate unique studies into human physiology and disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.

ABSTRACT
Induced pluripotent stem cells (iPSCs) are potentially valuable cell sources for disease models and future therapeutic applications; however, inefficient generation and the presence of integrated transgenes remain as problems limiting their current use. Here, we developed a new Sendai virus vector, TS12KOS, which has improved efficiency, does not integrate into the cellular DNA, and can be easily eliminated. TS12KOS carries KLF4, OCT3/4, and SOX2 in a single vector and can easily generate iPSCs from human blood cells. Using TS12KOS, we established iPSC lines from chimpanzee blood, and used DNA array analysis to show that the global gene-expression pattern of chimpanzee iPSCs is similar to those of human embryonic stem cell and iPSC lines. These results demonstrated that our new vector is useful for generating iPSCs from the blood cells of both human and chimpanzee. In addition, the chimpanzee iPSCs are expected to facilitate unique studies into human physiology and disease.

Show MeSH
Related in: MedlinePlus