Reprogramming induced pluripotent stem cells in the absence of c-Myc for differentiation into hepatocyte-like cells

Poly(ADP-ribose) polymerase 1 regulates nuclear reprogramming and promotes iPSC generation without c-Myc

“…Mouse iPSCs were gen erated from MEFs derived from 13.5 d old embryos of C57/B6 mice. The iPSCs were reprogrammed by the transduction of retroviral vectors encoding four transcription factors (Oct 4/Sox2/Klf4/c Myc; OSKM), or three tran scription factors (Oct 4/Sox2/Klf4; OSK), as described previously (Li et al, 2011). A total of 12 clones (Re 1 to Re 12; OSKM) were selected and estab lished.…”

“…For EB forma tion, iPSCs were dissociated into a single cell suspension by 0.25% trypsin EDTA and plated onto nonadherent culture dishes in DMEM with 15% FBS, 100 mM MEM nonessential amino acids, 0.55 mM 2 mercaptoethanol, and antibiotics at a density of 2 × 10 6 cells/100 mm plate. After 4 d in floating culture, EBs were transferred onto gelatin coated plates and maintained in the same medium for 24 h. EBs were then assigned for in vitro differentiation into tridermal lineages as previously described (Li et al, 2011). ALP activity, Alizarin red, and PAS staining.…”

“…Western blotting was performed as previously described (Li et al, 2011). The primary antibodies are listed in Table S4.…”

Krüppel-like factors in mammalian stem cells and development

“…Indeed, the Oct3/4, Sox2, Klf4 and c-Myc (OSKM) 'cocktail' or variations of it in multiple tissues of murine and human origin (fetal, neonatal and adult fibroblasts, primary hepatocytes, adult neuronal stem cells, menstrual blood stromal cell, prostate, bladder and ureter cells) can reprogram cells into iPSCs, with possible further differentiation into specialized cells such as gastric epithelial cells, lymphocytes, erythrocytes, myeloid cells, hepatocyte-like cells, retinal cells and melanocytes Kim et al, 2008;Kitada et al, 2012;Li et al, 2011;Park et al, 2008;Seiler et al, 2011;Takahashi et al, 2007;Takahashi and Yamanaka, 2006;Yang et al, 2011). As mentioned above, KLF4 has been identified as a factor that can reprogram mouse and human fibroblasts to induce their pluripotent state (Takahashi et al, 2007;Takahashi and Yamanaka, 2006).…”

Engrafted human stem cell–derived hepatocytes establish an infectious HCV murine model

“…In this context, the engraftment and maintenance of HLCs in a native liver parenchyma constitutes a relevant approach to promote further maturation and long-term residence of engrafted cells. Engraftment of human stem cell-derived HLCs has been described in different models of immunodeficient mice with transgene-induced (9,10,12) or chemically induced (9,(13)(14)(15)(16) liver toxicity with low efficiencies (12,14). Furthermore, not much is known about the effect of in situ engraftment on the level of maturation and permissiveness of these HLCs to infection by hepatitis viruses.…”