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Reprogramming Primary Amniotic Fluid and Membrane Cells to Pluripotency in Xeno-free Conditions
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1Mitchell Cancer Institute,University of South Alabama, 2College of Medicine,University of South Alabama, 3Institute for Regenerative Medicine,University of Zurich, 4Department of Dermatology,University Hospital Zurich, 5Center for Applied Biotechnology and Molecular Medicine (CABMM),University of Zurich – Irchel Campus
Chapters
- 00:05Title
- 00:50Isolation of Primary Mesenchymal Stem Cells from Human Amnion
- 02:31Harvesting Cultured Amniotic Fluid Stem Cells and Amniotic Membrane Stem Cells
- 03:47Transfection with Reprogramming Plasmids
- 05:35Picking Fully Reprogrammed Colonies for Clonial Expansion
- 06:43Clonal Expansion and Maturation of iPSCs
- 07:47Results: Morphological Progression of Cells During Reprogramming
- 08:43Conclusion
This protocol describes the reprogramming of primary amniotic fluid and membrane mesenchymal stem cells into induced pluripotent stem cells using a non-integrating episomal approach in fully chemically defined conditions. Procedures of extraction, culture, reprogramming, and characterization of the resulting induced pluripotent stem cells by stringent methods are detailed.
Tags
Keyword Extraction:
Amniotic FluidMembrane CellsPluripotencyXeno-free ConditionsInduced Pluripotent Stem CellsEpisomal ReprogrammingFetal Stem CellsDisease ModelingBasic ResearchLongitudinal StudiesChemically Defined ConditionsPediatric Cell-based TherapiesPlacentasAmnionRPMI-1640 MediumAFMC MediumAmniotic Fluid Stem CellsMembrane Stem CellsCell Detachment EnzymePBS
