Generation of a Human Induced Pluripotent Stem Cell Line Expressing a Magnetic Resonance Imaging Reporter Gene.
Yura SonPengsheng LiDakota OrtegaHuiliang QiuHannah PrachylMing YangWuqiang ZhuPublished in: Small methods (2024)
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.
Keyphrases
- magnetic resonance imaging
- contrast enhanced
- induced pluripotent stem cells
- endothelial cells
- magnetic resonance
- diffusion weighted imaging
- induced apoptosis
- oxidative stress
- gene expression
- machine learning
- crispr cas
- deep learning
- optical coherence tomography
- bone marrow
- genome editing
- big data
- endoplasmic reticulum stress
- genome wide identification