Direct reprogramming of somatic cells with the transcription factors Oct4 (also called Pou5f1), Sox2, Klf4 and Myc1 yields induced Pluripotent Stem (iPS) cells with marked similarity to Embryonic Stem (ES) cells. When the source of the cells is patients affected by various disorders iPS cells promise to be excellent models to study the gradual pathogenesis of disease. However, there is recent evidence that iPS cells derived from adult tissues retain residual DNA methylation signatures characteristic of their somatic tissue of origin, which appears to restrict their differentiation potential. Human amniotic fluid (AF) cells have been used for prenatal diagnosis of chromosomal and Mendelian disorders for decades. There is a subpopulation of cells in AF that expresses the tyrosine kinase receptor c-Kit appear to have stem cell-like properties. Developmentally and functionally, they represent a class of stem cells with intermediate characteristics between embryonic and adult stem cells. Our hypotheses are that c-kit expressing AF cells can be reprogrammed using fewer exogenous proteins than necessary with adult cells and that the resulting iPS cells will have methylation patterns more similar to embryonic stem cells than to adult derived iPS cells. Such pluripotent stem cell lines would provide models with which to investigate the underlying mechanisms of a variety of genetic diseases, leading to new diagnostic and treatment modalities. Human amniotic fluid may provide a source of cells capable of generating more applicable disease-specific iPS cells. Our specific aims are: 1) characterize subpopulations of cells obtained from AF after cytogenetic diagnosis has confirmed the presence of genetic abnormalities;2) determine minimal requirements for reprogramming AF cell subpopulations identified in Aim 1 to generate iPS cells. Our long-term goal is to develop banks of AF derived iPS cell lines representing different genetically related diseases.
Public Health Relevance: We propose to create stem cell lines for the study of genetic diseases from amniotic fluid cells left over after diagnosis. Such stem cell lines would provide excellent models with which to investigate the underlying mechanisms of a variety of genetic diseases, leading to new diagnostic and treatment modalities.
Thesaurus Terms: 21+ Years Old;Adult;Adult Human;Affect;Amniotic Fluid;Antenatal Diagnosis;Aqua Amnii;Biosynthetic Proteins;Blastocytes;Body Tissues;Cd117 Antigens;Cell Line;Cell Surface;Cellline;Cells;Characteristics;Chromosomal, Gene, Or Protein Abnormality;Cytogenetic Or Molecular Genetic Abnormality;Cytogenetics;Dna Methylation;Development;Diagnosis;Disease;Disease Model;Disorder;Es Cell;Embryo;Embryonic;Embryonic Cell;Exogenous Factors;Fetus;Genetic Abnormality;Genetic Condition;Genetic Diseases;Goals;Hereditary Disease;High-Risk Pregnancy;Human;Intrauterine Diagnosis;Loinc Axis 2 Property;Loinc Axis 4 System;Left;Liquor Amnii;Man (Taxonomy);Mast Cell Growth Factor Receptor;Methylation;Modality;Modeling;Modern Man;Molecular Abnormality;Molecular Disease;Nuclear;Ptk Receptors;Pathogenesis;Patients;Pattern;Pluripotent Stem Cells;Population;Prenatal Diagnosis;Principal Investigator;Progenitor Cells;Property;Protein Methylation;Proteins;Proto-Oncogene Protein C-Kit;Protocol;Protocols Documentation;Receptor Protein-Tyrosine Kinases;Recombinant Proteins;Regenerative Medicine;Residual;Residual State;Scf Receptor;Services;Somatic Cell;Source;Stem Cell Factor Receptor;Stem Cell Research;Stem Cells;Strains Cell Lines;System;Tissues;Transmembrane Receptor Protein Tyrosine Kinase;Tyrosine Kinase Growth Factor Receptor;Tyrosine Kinase Linked Receptors;Tyrosine Kinase Receptors;Undifferentiated;Validation;Viral Vector;Waters (Amniotic Fluid);Adult Human (21+);Adult Stem Cell;Adulthood;Antepartum Diagnosis;Blastomere Structure;C Kit;C-Kit Protein;C-Kit Receptor;Cell Bank;Cell Type;Cultured Cell Line;Developmental;Disease/Disorder;Disorder Model;Embryonic Stem Cell;Gene Product;Genetic Disorder;Hereditary Disorder;Human Disease;Ips;Induced Pluripotent Stem Cell;Kit Proto-Oncogene Protein;New Diagnostics;Next Generation Diagnostics;Novel Diagnostics;P145(C-Kit);P145c-Kit;Pluripotency;Programs;Small Molecule;Stem;Stem Cell Of Embryonic Origin;Transcription Factor;Transduction Efficiency
