Highly efficient mRNA-based gene transfer in feeder-free cultured H9 human embryonic stem cells

Peter Ponsaerts; Sjaak van der Sar; Viggo F I Van Tendeloo; Philippe G Jorens; Zwi N Berneman; Prim B Singh

doi:10.1089/clo.2004.6.211

Highly efficient mRNA-based gene transfer in feeder-free cultured H9 human embryonic stem cells

Peter Ponsaerts, Sjaak van der Sar, Viggo F I Van Tendeloo, Philippe G Jorens, Zwi N Berneman, Prim B Singh

Department of Medicine

Research output: Contribution to journal › Article › peer-review

19 Citations (Scopus)

Abstract

Several protocols have been described for virus-based gene transfer in human embryonic stem (hES) cells, while efficient non-viral methods are currently non-existing. In this study, we investigated the efficiency of mRNA-based gene transfer in feeder-free cultured H9 hES cells, based on electroporation of in vitro transcribed mRNA encoding the enhanced green fluorescent protein (EGFP). Optimisation of culture and electroporation conditions for feeder-free cultured H9 hES cells resulted a highly pure, transgene-expressing (90% positive cells) H9 hES cell population.

Original language	English
Pages (from-to)	211-6
Number of pages	6
Journal	Cellular Reprogramming
Volume	6
Issue number	3
DOIs	https://doi.org/10.1089/clo.2004.6.211
Publication status	Published - 2004

Keywords

Cell Survival
Cells, Cultured
Electroporation
Flow Cytometry
Green Fluorescent Proteins
Humans
Stem Cells
Transfection
Journal Article
Research Support, Non-U.S. Gov't

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title = "Highly efficient mRNA-based gene transfer in feeder-free cultured H9 human embryonic stem cells",

abstract = "Several protocols have been described for virus-based gene transfer in human embryonic stem (hES) cells, while efficient non-viral methods are currently non-existing. In this study, we investigated the efficiency of mRNA-based gene transfer in feeder-free cultured H9 hES cells, based on electroporation of in vitro transcribed mRNA encoding the enhanced green fluorescent protein (EGFP). Optimisation of culture and electroporation conditions for feeder-free cultured H9 hES cells resulted a highly pure, transgene-expressing (90% positive cells) H9 hES cell population.",

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language = "English",

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AU - Ponsaerts, Peter

AU - van der Sar, Sjaak

AU - Van Tendeloo, Viggo F I

AU - Jorens, Philippe G

AU - Berneman, Zwi N

AU - Singh, Prim B

PY - 2004

Y1 - 2004

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AB - Several protocols have been described for virus-based gene transfer in human embryonic stem (hES) cells, while efficient non-viral methods are currently non-existing. In this study, we investigated the efficiency of mRNA-based gene transfer in feeder-free cultured H9 hES cells, based on electroporation of in vitro transcribed mRNA encoding the enhanced green fluorescent protein (EGFP). Optimisation of culture and electroporation conditions for feeder-free cultured H9 hES cells resulted a highly pure, transgene-expressing (90% positive cells) H9 hES cell population.

KW - Cell Survival

KW - Cells, Cultured

KW - Electroporation

KW - Flow Cytometry

KW - Green Fluorescent Proteins

KW - Humans

KW - Stem Cells

KW - Transfection

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

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DO - 10.1089/clo.2004.6.211

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VL - 6

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JO - Cellular Reprogramming

JF - Cellular Reprogramming

SN - 2152-4971

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