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Small Methods, May 06 2024, | https://doi.org/10.1002/smtd.202301764
Generation of a Human Induced Pluripotent Stem Cell Line Expressing a Magnetic Resonance Imaging Reporter Gene
Yura Son 1, Pengsheng Li 1, Dakota Ortega 2, Huiliang Qiu 1, Hannah Prachyl 1, Ming Yang 3, Wuqiang Zhu 1
1Department of Cardiovascular Medicine, Department of Physiology and Biomedical Engineering, and Center for Regenerative Biotherapeutics, Mayo Clinic Arizona, Scottsdale, AZ, 85259, USA.
2KE Biosciences Preclinical Imaging Center, Arizona State University, Tempe, AZ, 85281, USA.
3Department of Radiology, Mayo Clinic Arizona, Scottsdale, AZ, 85259, USA.
CORRESPONDING AUTHOR : Wuqiang Zhu
Abstract
The objective of the current study is to develop a new method for tracking transplanted human induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs) using magnetic resonance imaging (MRI). The CRISPR/dCas9 activation system is employed to overexpress ferritin heavy chain (FHC) in hiPSC-CMs. The mRNA and protein expression of FHC in hiPSC and hiPSC-CMs significantly increased after transfection. Iron chloride does not affect the cell viability in a concentration range from 0 to 2000 µm. hiPSCs overexpressing FHC (hiPSC- FHC
OE) and hiPSC-CMs overexpressing FHC (hiPSC-CM-FHC
OE) significantly enhanced cellular uptake of iron chloride but with no changes in electrophysiological properties compared to hiPSC-CM-Control. Furthermore, hiPSC-CM-FHC
OE presented robust contrast and lower T
2* values, signifying their potential as highly effective candidates for cardiac MRI. Next, hiPSC-CM-FHC
OE is injected into mouse hearts and after 3 days of transplantation, MR images are obtained. hiPSC-CM-FHC
OE cells exhibited clear signals in the hearts with lower T
2* and rapid signal decay. Collectively, data from this proof-of-concept study demonstrated that endogenous labeling with FHC in hiPSC-CMs can be a potent strategy for enhancing the accuracy of cardiac MRI. This technology represents a significant step forward in tracking the transplanted hiPSC-CMs in the hearts of live animals.
논문정보
- Research article
- 2024년 05월 (BRIC 등록일 2024-05-08)
- 국외 연구진
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