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From Keratinocytes by Simple Transfection of Episomal Vectors Efficient Generation of Integration-Free Human Induced Pluripotent Stem Cells

SANDY SHEN,CHI HUNG, SHIANG, Y., LIM,RAYMOND CHING, BONG WONG, MINORU, H. S., KOa

semanticscholar(2014)

Cited 43|Views3
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Abstract
Keratinocytes represent an easily accessible cell source for derivation of human induced pluripotent stem (hiPS) cells, reportedly achieving higher reprogramming efficiency than fibroblasts. However, most studies utilized a retroviral or lentiviral method for reprogramming of keratinocytes, which introduces undesirable transgene integrations into the host genome. Moreover, current protocols of generating integration-free hiPS cells from keratinocytes are mostly inefficient. In this paper, we describe a more efficient, simple-to-use, and cost-effective method for generating integrationfree hiPS cells from keratinocytes. Our improved method using lipid-mediated transfection achieved a reprogramming efficiency of ∼0.14% on average. Keratinocyte-derived hiPS cells showed no integration of episomal vectors, expressed stem cell-specific markers and possessed potentials to differentiate into all three germ layers by in vitro embryoid body formation as well as in vivo teratoma formation. To our knowledge, this represents the most efficient method to generate integrationfree hiPS cells from keratinocytes. STEM CELLS TRANSLATIONAL MEDICINE 2014;3:787–791
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