Bottom: action potential duration at 90 and 50% repolarization (APD90 and APD50). (G) Top: maximal diastolic potential (MDP). Data correspond to clone UC C0406-iPS-C4P17 (also in G through I) similar results were observed with UC C0406-iPS-C1P17. (F) Representative spontaneous action potential tracing recorded by whole-cell patch clamp technique. ASGPR stands for asialoglycoprotein receptor. Glycogen accumulation was detected with periodic acid-Schiff staining. (D and E) Phase contrast and immunofluorescence photographs of hepatocytes and cardiomyocytes produced from representative UiPSC clones scale bars, 50 μm (phase contrast of cardiomyocytes), 200 μm (phase contrast of hepatocytes) and 50 μm (all immunofluorescence photographs). Bottom: confocal immunofluorescence of neurons and astrocytes (glial fibrillary acidic protein ) produced from the same UiPSC clone scale bars, 50 μm. Middle: confocal immunofluorescence microscopy for the indicated markers of neural rosettes produced from the same UiPSC clone scale bars, 50 μm. (C) Top, from left to right: phase contrast photographs of EBs growing in suspension, neural rosettes, and neurons produced from a representative UiPSC clone scale bars, 50 μm. AFP stands for α-fetoprotein scale bars, 50 μm. (B) Confocal immunofluorescence microscopy for markers of the 3 germ layers in differentiating embryoid bodies (EBs) of a representative UiPSC clone. (A) Teratomas comprising derivatives of the 3 germ layers. Multidifferentiation potential of urinary induced pluripotent stem cells (UiPSCs). (J) Bisulfite sequencing analysis for the Oct4 and Nanog proximal promoters in 2 representative UiPSC clones from the same donor. (I) Normal karyotype of representative male and female UiPSC clones. (H) Semiquantitative PCR showing integration of the exogenous transgenes in the genome of the indicated UiPSC clones, urine donor cells, water, and H9 ESCs were included as controls. (G) Scatter plot of DNA microarrays data for 2 representative UiPSC clones and H9 ESCs. (F) qPCR showing silencing of the exogenous transgenes in the indicated UiPSC clones values are referred to transduced cells extracted at day 6. Values are referred to donor urine cells H9 ESCs were used as control. hTERT indicates human telomerase reverse transcriptase. (E) Quantitative real-time PCR (qPCR) for endogenous ESC transcription factors in the indicated UiPSCs. (D) Confocal immunofluorescence microscopy for the indicated human embryonic stem cell (ESC) markers of a representative UiPSC clone scale bars, 100 μm. Bottom: representative phase contrast photographs of passage (P) 1 UiPSCs grown on feeders (alkaline phosphatase staining is also included) or Matrigel. (C) Top: representative phase contrast photographs of emerging urinary induced pluripotent stem cells (UiPSCs) colonies at different time points. Cells infected with the exogenous factors are also included note the early morphologic changes (clustering) indicative of reprogramming. (B) Phase contrast and immunofluorescence photographs of urine cells at day 3 after infection with control green fluorescent protein (GFP) retrovirus. SKOM refers to the four exogenous factors Sox2, Klf4, Oct4, and c-Myc. (A) Schematic representation of iPSC generation from urine cells (UC). Amander Clark and Insoo Hyun will consider the scientific, ethical, and policy issues related to IVG as a next generation approach for human reproduction.Generation of induced pluripotent stem cells (iPSCs) from urine cells. Animal models have successfully used IVG to restore fertility, and basic science research is currently translating this technology to human cells - with particular emphasis on reprogramming skin cells into stem cells, followed by IVG into gametes. IVG involves making gametes outside of the body either from stalled gamete precursor cells, or from skin cells that can be turned into egg or sperm cells in the laboratory. Instead, a new experimental procedure known as in vitro gametogenesis (IVG) could be used in these cases. However, IVF is not an option for persons who no longer make gametes. For persons who want a biologically related child, one of the most common treatments involves invitro fertilization (IVF). Infertility is a disease that affects millions of reproductive age men, women, and non-binary persons in the U.S. Department of Molecular, Cell and Developmental BiologyĬase Western Reserve University School of Medicine
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