Non-genetic Purification of Ventricular Cardiomyocytes from Differentiating Embryonic Stem Cells through Molecular Beacons Targeting IRX-4

Kiwon Ban, Brian Wile, Kyu Won Cho, Sangsung Kim, Ming Ke Song, Sang Yoon Kim, Jason Singer, Anum Syed, Shan Ping Yu, Mary Wagner, Gang Bao, Young Sup Yoon

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Isolation of ventricular cardiomyocytes (vCMs) has been challenging due to the lack of specific surface markers. Here we show that vCMs can be purified from differentiating mouse embryonic stem cells (mESCs) using molecular beacons (MBs) targeting specific intracellular mRNAs. We designed MBs (IRX4 MBs) to target mRNA encoding Iroquois homeobox protein 4 (Irx4), a transcription factor specific for vCMs. To purify mESC vCMs, IRX4 MBs were delivered into cardiomyogenically differentiating mESCs, and IRX4 MBs-positive cells were FACS-sorted. We found that, of the cells isolated, ∼98% displayed vCM-like action potentials by electrophysiological analyses. These MB-purified vCMs continuously maintained their CM characteristics as verified by spontaneous beating, Ca2+ transient, and expression of vCM-specific proteins. Our study shows the feasibility of isolating pure vCMs via cell sorting without modifying host genes. The homogeneous and functional ventricular CMs generated via the MB-based method can be useful for disease investigation, drug discovery, and cell-based therapies.

Original languageEnglish
Pages (from-to)1239-1249
Number of pages11
JournalStem Cell Reports
Volume5
Issue number6
DOIs
Publication statusPublished - 2015 Dec 8

Fingerprint

Embryonic Stem Cells
Stem cells
Cardiac Myocytes
Purification
Cells
Homeodomain Proteins
Messenger RNA
Sorting
Transcription Factors
Genes
Feasibility Studies
Drug Discovery
Cell- and Tissue-Based Therapy
Action Potentials
Proteins
Mouse Embryonic Stem Cells

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Genetics
  • Developmental Biology
  • Cell Biology

Cite this

Ban, Kiwon ; Wile, Brian ; Cho, Kyu Won ; Kim, Sangsung ; Song, Ming Ke ; Kim, Sang Yoon ; Singer, Jason ; Syed, Anum ; Yu, Shan Ping ; Wagner, Mary ; Bao, Gang ; Yoon, Young Sup. / Non-genetic Purification of Ventricular Cardiomyocytes from Differentiating Embryonic Stem Cells through Molecular Beacons Targeting IRX-4. In: Stem Cell Reports. 2015 ; Vol. 5, No. 6. pp. 1239-1249.
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Ban, K, Wile, B, Cho, KW, Kim, S, Song, MK, Kim, SY, Singer, J, Syed, A, Yu, SP, Wagner, M, Bao, G & Yoon, YS 2015, 'Non-genetic Purification of Ventricular Cardiomyocytes from Differentiating Embryonic Stem Cells through Molecular Beacons Targeting IRX-4', Stem Cell Reports, vol. 5, no. 6, pp. 1239-1249. https://doi.org/10.1016/j.stemcr.2015.10.021

Non-genetic Purification of Ventricular Cardiomyocytes from Differentiating Embryonic Stem Cells through Molecular Beacons Targeting IRX-4. / Ban, Kiwon; Wile, Brian; Cho, Kyu Won; Kim, Sangsung; Song, Ming Ke; Kim, Sang Yoon; Singer, Jason; Syed, Anum; Yu, Shan Ping; Wagner, Mary; Bao, Gang; Yoon, Young Sup.

In: Stem Cell Reports, Vol. 5, No. 6, 08.12.2015, p. 1239-1249.

Research output: Contribution to journalArticle

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AU - Ban, Kiwon

AU - Wile, Brian

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AU - Kim, Sangsung

AU - Song, Ming Ke

AU - Kim, Sang Yoon

AU - Singer, Jason

AU - Syed, Anum

AU - Yu, Shan Ping

AU - Wagner, Mary

AU - Bao, Gang

AU - Yoon, Young Sup

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