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 language | English |
|---|---|
| Pages (from-to) | 1239-1249 |
| Number of pages | 11 |
| Journal | Stem Cell Reports |
| Volume | 5 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - 2015 Dec 8 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Genetics
- Developmental Biology
- Cell Biology
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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 journal › Article
TY - JOUR
T1 - Non-genetic Purification of Ventricular Cardiomyocytes from Differentiating Embryonic Stem Cells through Molecular Beacons Targeting IRX-4
AU - Ban, Kiwon
AU - Wile, Brian
AU - Cho, Kyu Won
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
PY - 2015/12/8
Y1 - 2015/12/8
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84949488041&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84949488041&partnerID=8YFLogxK
U2 - 10.1016/j.stemcr.2015.10.021
DO - 10.1016/j.stemcr.2015.10.021
M3 - Article
C2 - 26651608
AN - SCOPUS:84949488041
VL - 5
SP - 1239
EP - 1249
JO - Stem Cell Reports
JF - Stem Cell Reports
SN - 2213-6711
IS - 6
ER -