Journal: Proceedings of the Japan Academy. Series B, Physical and Biological Sciences

Article Title: Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome

doi: 10.2183/pjab.85.348

Figure Lengend Snippet: Expression of exogenous genes in human fibroblasts by SeV vectors. A. Efficient induction of GFP cDNA by TSΔF/SeV in BJ and HDF at an MOI of 3. BC: bright contrast. B. Schematic presentation of SeV vector genomes. Reprogramming genes were inserted at 18+, PM, HN, HNL and Leis (L), respectively. The expression levels of inserted genes decreased depending on the inserted site (polar effect: Refs. ) as shown by Western blotting on day 3 after infection. Anti-SeV blot was performed to confirm equal infection efficiency of the vectors.

Article Snippet: Human fibroblast BJ from neonatal foreskin (ATCC, USA), and HDF from facial dermis of 36-year femail (Cell Applications, Inc., USA), were maintained in fibroblast growth medium (Cell Applications, Inc., USA) or DMEM (Invitrogen, USA) supplemented with 10% FBS.

Techniques: Expressing, Plasmid Preparation, Western Blot, Infection

Journal: Proceedings of the Japan Academy. Series B, Physical and Biological Sciences

Article Title: Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome

doi: 10.2183/pjab.85.348

Figure Lengend Snippet: Genomic Southern blot, karyotyping and fingerprinting of SeV-iPSC. A. DNA fingerprinting of SeV-iPS clones. PCR analysis of three variable number of tandem repeats (VNTR) loci of D17S1290, MCT118 and ApoB-100 using genomic DNA from the SeV-iPS clones confirmed that these clones were originated from human fibroblasts BJ or HDF. B1, HNL1 and HNL5 were derived from BJ; XH1, 7H5, 7H8 and 7H10 were from HDF. B. Viral transgenes were not detected from the host genome as analyzed by genomic Southern blot. C. Karyotyping of SeV-iPSC. Viral-free SeV-iPSC HNLs at passage 34 were used for karyotyping.

Article Snippet: Human fibroblast BJ from neonatal foreskin (ATCC, USA), and HDF from facial dermis of 36-year femail (Cell Applications, Inc., USA), were maintained in fibroblast growth medium (Cell Applications, Inc., USA) or DMEM (Invitrogen, USA) supplemented with 10% FBS.

Techniques: Southern Blot, DNA Profiling, Clone Assay, Derivative Assay

Journal: Proceedings of the Japan Academy. Series B, Physical and Biological Sciences

Article Title: Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome

doi: 10.2183/pjab.85.348

Figure Lengend Snippet: In vivo pluripotency of transgenes-free SeV-iPSC. Hematoxylin and eosin staining of teratoma sections of SeV-iPS clones (6 weeks post-injection into SCID mice). Tissues were differentiated from Tg-free human neonatal fibroblast BJ-derived HNLs (A to C), HNL1 (D to I), and adult fibroblast HDF-derived XH1 (J to L) containing multiple tissues derived from three germ layers: glandular structures (A, G, K), cartilage (B, J), bone (C, F, white arrows) and bone marrow-like structure (F), epithelium (J, D), transitional epithelium (E), population of secreting-like cells (B, D, indicated by black arrows), muscle (C, I, K, L), and glomerulus of kidney-like tissue (H).

Article Snippet: Human fibroblast BJ from neonatal foreskin (ATCC, USA), and HDF from facial dermis of 36-year femail (Cell Applications, Inc., USA), were maintained in fibroblast growth medium (Cell Applications, Inc., USA) or DMEM (Invitrogen, USA) supplemented with 10% FBS.

Techniques: In Vivo, Staining, Clone Assay, Injection, Derivative Assay