Disease-Specific Induced Pluripotent Stem Cells.

“In contrast, introducing a set of transcription factors linked to pluripotency can directly reprogram human somatic cells to produce induced pluripotent stem (iPS) cells, a method that has been achieved by several groups worldwide (Lowry et al., 2008; Park et al., 2008b; Takahashi et al., 2007; Yu et al., 2007).”

“In contrast, introducing a set of transcription factors linked to pluripotency can directly reprogram human somatic cells to produce induced pluripotent stem (iPS) cells, a method that has been achieved by several groups worldwide (Lowry et al., 2008; Park et al., 2008b; Takahashi et al., 2007; Yu et al., 2007).”

“Following 2 to 3 weeks of culture in hES cell-supporting conditions, compact refractile ES-like colonies emerged among a background of fibroblasts, as previously described (Park et al., 2008a, 2008b).”

“Strategies for inducing specific mutation or gene repair by homologous recombination were first established for the HPRT locus (Doetschman et al., 1987, 1988; Thomas and Capecchi, 1987).”

“Human embryonic stem cells isolated from excess embryos from in vitro fertilization clinics represent an immortal propagation of pluripotent cells that theoretically can generate any cell type within the human body (Lerou et al., 2008; Murry and Keller, 2008).”

“Multiplex PCR analysis with primer sets amplifying several (but not all) intragenic intervals of the dystrophin gene (Beggs et al., 1990; Chamberlain et al., 1988) revealed the deletion of exons 45–52 in the iPS cells derived from a patient with Duchenne muscular dystrophy (DMD; Figure 1C).”

“In contrast, introducing a set of transcription factors linked to pluripotency can directly reprogram human somatic cells to produce induced pluripotent stem (iPS) cells, a method that has been achieved by several groups worldwide (Lowry et al., 2008; Park et al., 2008b; Takahashi et al., 2007; Yu et al., 2007).”

“…from a carrier of Lesch-Nyhan syndrome using five doxycycline-inducible lentiviral vectors (OCT4, SOX2, KLF4, c-MYC, and NANOG), a strategy that has been used to isolate murine iPS cells (Brambrink et al., 2008; Stadtfeld et al., 2008) but previously had not been attempted with human somatic cells.”

“…from a carrier of Lesch-Nyhan syndrome using five doxycycline-inducible lentiviral vectors (OCT4, SOX2, KLF4, c-MYC, and NANOG), a strategy that has been used to isolate murine iPS cells (Brambrink et al., 2008; Stadtfeld et al., 2008) but previously had not been attempted with human somatic cells.”

“Strategies for inducing specific mutation or gene repair by homologous recombination were first established for the HPRT locus (Doetschman et al., 1987, 1988; Thomas and Capecchi, 1987).”

“…Building 7214, 300 Longwood Avenue, Boston, MA 02115 2Massachusetts General Hospital Cancer Center and Center for Regenerative Medicine and Department of Stem Cell and Regenerative Biology, 185 Cambridge Street, Boston, MA 02114, USA 3Department of Molecular and Cellular Biology, Harvard…”

“In a proof of principle experiment, NT-embryonic stem (ES) cells generated from mice with genetic immunodeficiency were used to combine gene and cell therapy to repair the genetic defect (Rideout et al., 2002).”

“Some studies place the frequency of spontaneous fetal demise (miscarriage) in DS to be above 40% (Bittles et al., 2007).”

“Following 2 to 3 weeks of culture in hES cell-supporting conditions, compact refractile ES-like colonies emerged among a background of fibroblasts, as previously described (Park et al., 2008a, 2008b).”

“Multiplex PCR analysis with primer sets amplifying several (but not all) intragenic intervals of the dystrophin gene (Beggs et al., 1990; Chamberlain et al., 1988) revealed the deletion of exons 45–52 in the iPS cells derived from a patient with Duchenne muscular dystrophy (DMD; Figure 1C).”

“A mouse model for the DS critical region on distal human chromosome 21 fails to recapitulate the human cranial abnormalities commonly associated with trisomy 21 (Olson et al., 2004).”

“Murine strains carrying the same genetic deficiencies as the human bone marrow failure disease Fanconi anemia demonstrate DNA repair defects consistent with the human condition (e.g., Chen et al., 1996), yet none develop the spontaneous bone marrow failure that is the hallmark of the human disease.”

“Strategies for inducing specific mutation or gene repair by homologous recombination were first established for the HPRT locus (Doetschman et al., 1987, 1988; Thomas and Capecchi, 1987).”

“The ultimate standard of pluripotency for human cells is teratoma formation in immunodeficient murine hosts (Lensch et al., 2007).”

“As BMD is a milder form of disease, and the dystrophin gene one of the largest in the human genome, definition of the genetic lesion responsible for this condition is sometimes elusive (Prior and Bridgeman, 2005).”

“Molecular analysis of the GBA gene in the Gaucher disease line revealed a 1226A > G point mutation, causing a N370S amino acid substitution (Figure 1B); the second allele is known to have a frameshifting insertion of a single guanine at cDNA nucleotide 84 (84GG) (Beutler et al., 1991).”

“…(Huntington disease; HD) and verified the presence of expanded (CAG)n polyglutamine triplet repeat sequences (72) in the proximal portion of the huntingtin gene (Figure 1C; Riess et al., 1993) in one allele and 19 repeats in the other (where the normal range is 35 or less; Chong et al., 1997).”

“The Lesch-Nyhan syndrome carrier line harbors heterozygous deficiency of the HPRT gene (Nussbaum et al., 1983).”

“Most of the human cell lines in wide use today carry genetic and epigenetic artifacts of accommodation to tissue culture and are derived either from malignant tissues or are genetically modified to drive immortal growth (Grimm, 2004).”

“Human embryonic stem cells isolated from excess embryos from in vitro fertilization clinics represent an immortal propagation of pluripotent cells that theoretically can generate any cell type within the human body (Lerou et al., 2008; Murry and Keller, 2008).”

“We generated iPS cell lines from a patient with Huntington chorea (Huntington disease; HD) and verified the presence of expanded (CAG)n polyglutamine triplet repeat sequences (72) in the proximal portion of the huntingtin gene (Figure 1C; Riess et al., 1993) in one allele and 19 repeats in the other (where the normal…”

“tions were performed using 50 ng of genomic DNA with primers corresponding to the mutated regions of genes responsible for each condition (ADA-SCID, GD, SBDS [ Calado et al., 2007 ], and HD).”

“…genetic diagnosis (PGD; genetic analysis of single blastomeres obtained by embryo biopsy) can yield stem cell lines that model single-gene disorders (Verlinsky et al., 2005), but the vast majority of diseases that show more complex genetic patterns of inheritance are not represented in this pool.”

“Orthologous segments to human chromosome 21 are present on mouse chromosomes 10 and 17, and distal human chromosome 21 corresponds to mouse chromosome 16 where trisomy 16 in the mouse is lethal (Nelson and Gibbs, 2004).”

“Bone marrow mesenchymal cells from SBDS patient (SBDS, DF250) have been described (Austin et al., 2005).”

“As reported previously with either lentiviral or Moloney-based retroviral vectors (Dimos et al., 2008; Ebert et al., 2009; Hockemeyer et al., 2008; Park et al., 2008a; Yu et al., 2007), we detected residual expression of the reprogramming factors for most of the transgenes in all cell lines with…”

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