Cell Reprogramming Methods and Protocols

This volume provides an understanding of the factors involved in nuclear reprogramming, which is essential for the success of reprogramming. The book is aimed at reprogramming differentiated cells and germ line transmission of pluripotent stem cells and features chapters that deal with reprogramming-related issues such as analysis of mitochondrial DNA in reprogrammed cells and the isolation of reprogramming intermediates; alternative methods for nuclear transfer; the production of germ-line chimeras from embryonic stem cells and induced pluripotent stem cells; and neonatal care and management of somatic cell nuclear transfer derived offspring. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, is the perfect guide for molecular biologists, stem cell biologists, clinicians, biotechnologists, students, veterinarians and animal care technicians involved with reprogramming, nuclear transfer and transgenesis.

Cellular Reprogramming in Basic and Applied Biomedicine: The Dawn of Regenerative Medicine.- Synthetic mRNA Reprogramming of Human Fibroblast Cells.- MicroRNA Mediated Reprogramming of Somatic Cells into Induced Pluripotent Stem Cells.- Generation of Footprint-Free Induced Pluripotent Stem Cells from Human Fibroblasts Using Episomal Plasmid Vectors.- Reprogramming of Human Fibroblasts with Non-Integrating RNA Virus on Feeder-Free or Xeno-Free Conditions.- Inducing Pluripotency in Cattle.- Generation of Induced Pluripotent Stem Cells (iPSCs) From Adult Canine Fibroblasts.- Derivation of Equine-Induced Pluripotent Stem Cell Lines Using a PiggyBac Transposon Delivery System and Temporal Control of Transgene Expression.- Generation of Induced Pluripotent Stem Cells from Mammalian Endangered Species.- Generation of Efficient Germ-Line Chimeras Using Embryonic Stem Cells.- Generation of Viable Mice from Induced Pluripotent Stem Cells (iPSCs) Through Tetraploid Complementation.- Cloning Endangered Felids by Interspecies Somatic Cell Nuclear Transfer.- Generation of Chimeras from Porcine Induced Pluripotent Stem Cells.- A Novel Method on Somatic Cell Nuclear Transfer with the Minimum Equipment.- Neonatal Care and Management of Foals Derived by Somatic Cell Nuclear Transfer.- Isolation of Reprogramming Intermediates During Generation of Induced Pluripotent Stem Cells from Mouse Embryonic Fibroblasts.- Analysis of Mitochondrial DNA in Induced Pluripotent and Embryonic Stem Cells.- Genome Modification of Pluripotent Cells By Using Transcription Activator-Like Effector Nucleases (TALENs).