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Curr Protoc Stem Cell Biol. 2018 May;45(1):e48. doi: 10.1002/cpsc.48. Epub 2018 May 4.

Generation of Human Induced Pluripotent Stem Cells Using a Defined, Feeder-Free Reprogramming System.

Author information

1
Center for Regenerative Medicine (CReM) and Section of Gastroenterology, Department of Medicine, Boston University School of Medicine, Boston, Massachusetts.

Abstract

Human induced pluripotent stem cells (hiPSCs) offer great opportunities for the study of human development and disease modeling and have enormous potential for use in future clinical cell-based therapies. However, most current systems to create hiPSCs often expose the cells to animal feeder layers or xenogeneic reagents; this raises safety concerns about using hiPSC-derived cells for therapeutic purposes. Here, we describe protocols to generate hiPSCs without exposing the cells to xenogeneic materials that uses a defined, feeder-free reprogramming system. With this method, we were able to successfully reprogram not only patient-derived peripheral blood mononuclear cells but also amniocytes from the amniotic fluid of stillborn fetuses using two independent reprogramming platforms. Importantly, hiPSCs generated in this fashion expressed pluripotent markers and had normal karyotypes. The protocols allowed us to generate and culture hiPSCs under Good Manufacturing Practice-like conditions, a necessary step for the future clinical application of these cells.

KEYWORDS:

STEMCCA lentiviral vector; Sendai virus vector; amniocytes; feeder-free reprogramming system; human induced pluripotent stem cells (hiPSCs); peripheral blood mononuclear cells (PBMCs)

PMID:
30040234
PMCID:
PMC6060628
DOI:
10.1002/cpsc.48
[Indexed for MEDLINE]
Free PMC Article

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