Abstract: The invention provides a method of preparing a nucleic acid population suitable for RNA sequencing. The method involves amplifying a double-stranded DNA and a poly T sequence by using the DNA constituted of any additional nucleic acid sequence X, poly T sequence, mRNA sequence isolated from a biological sample, poly A sequence and any additional nucleic acid sequence Y in this order as a template, a first primer containing any additional nucleic acid sequence X having amine added to the 5?-terminal (and a poly T sequence), and a second primer containing any additional nucleic acid sequence Y (and a poly T sequence), followed by fractionalizing the DNA, phosphorylating the DNA, preparing cDNA by using the DNA as a template and a third primer, adding adenine (A) to the cDNA, linking a DNA, and amplifying the DNA by using the DNA as a template, a fourth primer, and a fifth primer.

Abstract: The present invention provides a method for expanding PGC/PGCLC, including culturing PGC/PGCLC in the presence of a phosphodiesterase 4 (PDE4) inhibitor and/or cyclosporine A, further in the presence of forskolin, and a method for inducing oocytes from PGC/PGCLC, including culturing PGC/PGCLC in the presence of bone forming protein (BMP) and retinoic acid (RA).

Abstract: Provided are a method for producing a spermatogenic stem cell-like cell from a primordial germ cell-like cell derived from an isolated pluripotent stem cell in vitro, the method including (1) a step of coculturing a primordial germ cell-like cell with a gonad somatic cell in suspension to give reconstituted testis, and (2) a step of culturing the obtained reconstituted testis at gas/liquid interface to induce a DDX4-positive and PLZF-positive cell in the reconstituted testis; and a method for producing a GSC-like cell, including dissociating a spermatogenic stem cell-like cell obtained by the method from the reconstituted testis, and culturing the cell under conditions that can induce a germline stem cell from the spermatogenic stem cell.

Abstract: The invention provides a method for inducing human primordial germ cell-like (PGC-like) cells from human pluripotent stem cells, with high efficiency and high reproducibility, and a cell surface marker for identifying human PGC-like cells. In particular, the invention provides a method for producing a human PGC-like cell from a human pluripotent stem cell, includes a step of producing a mesoderm-like cell by culturing a human pluripotent stem cell in a culture medium comprising activin A and a GSK3? inhibitor, and a step of culturing the mesoderm-like cell in a culture medium containing BMP. The invention also provides a method for producing an isolated human PGC-like cell, which includes the aforementioned two steps and the additional step of selecting a cell positive to at least one cell surface marker selected from the group consisting of PECAM (CD31), INTEGRINa6 (CD49f), INTEGRIN?3 (CD61), KIT (CD117), EpCAM, PODOPLANIN and TRA1-81.

Abstract: The invention provides a method for inducing human primordial germ cell-like (PGC-like) cells from human pluripotent stem cells, with high efficiency and high reproducibility, and a cell surface marker for identifying human PGC-like cells. In particular, the invention provides a method for producing a human PGC-like cell from a human pluripotent stem cell, includes a step of producing a mesoderm-like cell by culturing a human pluripotent stem cell in a culture medium comprising activin A and a GSK3? inhibitor, and a step of culturing the mesoderm-like cell in a culture medium containing BMP. The invention also provides a method for producing an isolated human PGC-like cell, which includes the aforementioned two steps and the additional step of selecting a cell positive to at least one cell surface marker selected from the group consisting of PECAM (CD31), INTEGRIN?6 (CD49f), INTEGRIN?3 (CD61), KIT (CD117), EpCAM, PODOPLANIN and TRA1-81.

Abstract: This invention provides a method of producing a primordial germ cell-like cell (PGCLC) from an epiblast isolated from an embryo or an epiblast-like cell (EpiLC) induced from a pluripotent stem cell (PSC), which comprises allowing the epiblast or EpiLC to express exogenous transcription factor(s) selected from the group consisting of: (i) Blimp1, Prdm14 and Tfap2c; ii) Blimp1 and Prdm14; (iii) Blimp1 and Tfap2c; (iv) Prdm14 and Tfap2c; and (v) Prdm14; thereby inducing the epiblast or EpiLC into a PGC state without acquiring transient mesodermal program.

Abstract: The invention provides a method for inducing human primordial germ cell-like (PGC-like) cells from human pluripotent stem cells, with high efficiency and high reproducibility, and a cell surface marker for identifying human PGC-like cells. In particular, the invention provides a method for producing a human PGC-like cell from a human pluripotent stem cell, includes a step of producing a mesoderm-like cell by culturing a human pluripotent stem cell in a culture medium comprising activin A and a GSK3? inhibitor, and a step of culturing the mesoderm-like cell in a culture medium containing BMP. The invention also provides a method for producing an isolated human PGC-like cell, which includes the aforementioned two steps and the additional step of selecting a cell positive to at least one cell surface marker selected from the group consisting of PECAM (CD31), INTEGRIN?6 (CD49f), INTEGRIN?3 (CD61), KIT (CD117), EpCAM, PODOPLANIN and TRA1-81.

Abstract: This invention provides a method of producing an epiblast-like cell (EpiLC) from a pluripotent stem cell, which comprises culturing the pluripotent stem cell in the presence of activin A; a method of producing a primordial germ cell-like (PGC-like) cell a pluripotent stem cell, which comprises culturing the EpiLC obtained by the method above in the presence of BMP4 and LIF. Also provided are a cell population containing PGC-like cells as obtained by the method, and reagent kits for the EpiLC- and PGC-like cell-induction from a pluripotent stem cell.

Abstract: The invention provides a method of preparing a nucleic acid population suitable for RNA sequencing. The method involves amplifying a double-stranded DNA and a poly T sequence by using the DNA constituted of any additional nucleic acid sequence X, poly T sequence, mRNA sequence isolated from a biological sample, poly A sequence and any additional nucleic acid sequence Y in this order as a template, a first primer containing any additional nucleic acid sequence X having amine added to the 5?-terminal (and a poly T sequence), and a second primer containing any additional nucleic acid sequence Y (and a poly T sequence), followed by fractionalizing the DNA, phosphorylating the DNA, preparing cDNA by using the DNA as a template and a third primer, adding adenine (A) to the cDNA, linking a DNA, and amplifying the DNA by using the DNA as a template, a fourth primer, and a fifth primer.

Abstract: This invention provides a method of producing a primordial germ cell-like cell (PGCLC) from an epiblast isolated from an embryo or an epiblast-like cell (EpiLC) induced from a pluripotent stem cell (PSC), which comprises allowing the epiblast or EpiLC to express exogenous transcription factor(s) selected from the group consisting of: (i) Blimp1, Prdm14 and Tfap2c; (ii) Blimp1 and Prdm14; (iii) Blimp1 and Tfap2c; (iv) Prdm14 and Tfap2c; and (v) Prdm14; thereby inducing the epiblast or EpiLC into a PGC state without acquiring transient mesodermal program.

Abstract: A method for amplification of a nucleotide sequence characterized by performing PCR amplification using mRNA isolated from a biological sample as a template and using a first primer comprising a poly(T) sequence and an additional nucleotide sequence X thereto and a second primer comprising a poly(T) sequence and an additional nucleotide sequence Y thereto, provided that the nucleotide sequences X and Y in the first and second primers, respectively, have different sequences from each other.

Abstract: This invention provides a method of producing an epiblast-like cell (EpiLC) from a pluripotent stem cell, which comprises culturing the pluripotent stem cell in the presence of activin A; a method of producing a primordial germ cell-like (PGC-like) cell a pluripotent stem cell, which comprises culturing the EpiLC obtained by the method above in the presence of BMP4 and LIF. Also provided are a cell population containing PGC-like cells as obtained by the method, and reagent kits for the EpiLC- and PGC-like cell-induction from a pluripotent stem cell.

Abstract: We describe two primordial germ cell-specifically expressed genes, GCR1 (Fragilis) and GCR2 (Stella), as well as their fragments, homologues, variants or deriviatives thereof which are markers for primordial germ cells and may be used to identify such cells in cell populations.

Abstract: A method for amplification of a nucleotide sequence characterized by performing PCR amplification using mRNA isolated from a biological sample as a template and using a first primer comprising a poly(T) sequence and an additional nucleotide sequence X thereto and a second primer comprising a poly(T) sequence and an additional nucleotide sequence Y thereto, provided that the nucleotide sequences X and Y in the first and second primers, respectively, have different sequences from each other.

Abstract: The present invention relates to two primordial germ cell-specific expressed genes, Fragilis and Stella. The sequences and sues of human Stella and Fragilis are disclosed herein, as are several mouse sequences related to Fragilis. The present invention relates to the use of Stella and Fragilis as markers for primordial germ cells and can be used to identify such cells. Additionally, the present invention relates to the use of Stella and Fragilis for the diagnosis, treatment and/or prevention of disease.

Abstract: The present invention relates to two primordial germ cell-specific expressed genes, Fragilis and Stella. The sequences and sues of human Stella and Fragilis are disclosed herein, as are several mouse sequences related to Fragilis. The present invention relates to the use of Stella and Fragilis as markers for primordial germ cells and can be used to identify such cells. Additionally, the present invention relates to the use of Stella and Fragilis for the diagnosis, treatment and/or prevention of disease.

Abstract: We describe two primordial germ cell-specifically expressed genes, GCR1 (Fragilis) and GCR2 (Stella), as well as their fragments, homologues, variants or derivatives thereof which are markers for primordial germ cells and may be used to identify such cells in cell populations.

Abstract: We describe two primordial germ cell-specifically expressed genes, GCR1 (Fragilis) and GCR2 (Stella), as well as their fragments, homologues, variants or deriviatives thereof which are markers for primordial germ cells and may be used to identify such cells in cell populations.

Abstract: The present invention relates to two primordial germ cell-specific expressed genes, Fragilis and Stella. The sequences and sues of human Stella and Fragilis are disclosed herein, as are several mouse sequences related to Fragilis. The present invention relates to the use of Stella and Fragilis as markers for primordial germ cells and can be used to identifiy such cells. Additionally, the present inveniton relates to the use of Stella and Fragilis for the diagnosis, treatment and/or prevention of disease.

Abstract: The present invention describes and claims two primordial germ cell-specifically expressed genes, GCR1 (Fragilis) and GCR2 (Stella), which are markers for primordial germ cells and may be used to identify such cells in cell populations.