Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles.

PurposeThe delivery of transgenes into human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (hiPSC-CMs) represents an important tool in cardiac regeneration with potential for clinical applications. Gene transfection is more difficult, however, for hiPSCs and hiPSC-CMs than for somatic cells. Despite improvements in transfection and transduction, the efficiency, cytotoxicity, safety, and cost of these methods remain unsatisfactory. The objective of this study is to examine gene transfection in hiPSCs and hiPSC-CMs using magnetic nanoparticles (NPs).MethodsMagnetic NPs are unique transfection reagents that form complexes with nucleic acids by ionic interaction. The particles, loaded with nucleic acids, can be guided by a magnetic field to allow their concentration onto the surface of the cell membrane. Subsequent uptake of the loaded particles by the cells allows for high efficiency transfection of the cells with nucleic acids. We developed a new method using magnetic NPs to transfect hiPSCs and hiPSC-CMs. HiPSCs and hiPSC-CMs were cultured and analyzed using confocal microscopy, flow cytometry, and patch clamp recordings to quantify the transfection efficiency and cellular function.ResultsWe compared the transfection efficiency of hiPSCs with that of human embryonic kidney (HEK 293) cells. We observed that the average efficiency in hiPSCs was 43%±2% compared to 62%±4% in HEK 293 cells. Further analysis of the transfected hiPSCs showed that the differentiation of hiPSCs to hiPSC-CMs was not altered by NPs. Finally, robust transfection of hiPSC-CMs with an efficiency of 18%±2% was obtained.ConclusionThe difficult-to-transfect hiPSCs and hiPSC-CMs were efficiently transfected using magnetic NPs. Our study offers a novel approach for transfection of hiPSCs and hiPSC-CMs without the need for viral vector generation

High-Velocity Saturation in Graphene Encapsulated by Hexagonal Boron Nitride

We measure drift velocity in monolayer graphene encapsulated by hexagonal boron nitride (hBN), probing its dependence on carrier density and temperature. Due to the high mobility (>5 × 10⁴ cm²/V/s) of our samples, the drift velocity begins to saturate at low electric fields (∼0.1 V/μm) at room temperature. Comparing results to a canonical drift velocity model, we extract room-temperature electron saturation velocities ranging from 6 × 10⁷ cm/s at a low carrier density of 8 × 10¹¹ cm^–2 to 2.7 × 10⁷ cm/s at a higher density of 4.4 × 10¹² cm^–2. Such drift velocities are much higher than those in silicon (∼10⁷ cm/s) and in graphene on SiO₂, likely due to reduced carrier scattering with surface optical phonons whose energy in hBN (>100 meV) is higher than that in other substrates

Highly efficient transfection of human induced pluripotent stem cells using magnetic nanoparticles

The delivery of transgenes into human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (hi PSC-C Ms) represents an important tool in cardiac regeneration with potential for clinical applications. Gene transfection is more difficult, however, for hiPSCs and hi PSC-CMs than for somatic cells. Despite improvements in transfection and transduction, the efficiency, cytotoxicity, safety, and cost of these methods remain unsatisfactory. The objective of this study is to examine gene transfection in hiPSCs and hiPSC-CMs using magnetic nanoparticles (NPs). Methods: Magnetic NPs are unique transfection reagents that form complexes with nucleic acids by ionic interaction. The particles, loaded with nucleic acids, can be guided by a magnetic field to allow their concentration onto the surface of the cell membrane. Subsequent uptake of the loaded particles by the cells allows for high efficiency transfection of the cells with nucleic acids. We developed a new method using magnetic NPs to transfect hiPSCs and hiPSC-CMs. HiPSCs and hiPSC-CMs were cultured and analyzed using confbcal microscopy, flow cytometry, and patch clamp recordings to quantify the transfection efficiency and cellular function. Results: We compared the transfection efficiency of hiPSCs with that of human embryonic kidney (HEK 293) cells. We observed that the average efficiency in hi PSCs was 43%+/- 2% compared to 62%+/- 4% in HEK 293 cells. Further analysis of the transfected hiPSCs showed that the differentiation of hiPSCs to hi PSC-CMs was not altered by NPs. Finally, robust transfection of hi PSC-CMs with an efficiency of I8%+/- 2% was obtained. Conclusion: The difficult-to-transfect hiPSCs and hiPSC-CMs were efficiently transfected using magnetic NPs. Our study offers a novel approach for transfection of hiPSCs and hiPSCCMs without the need for viral vector generation

Hexagonal boron nitride as a low-loss dielectric for superconducting quantum circuits and qubits

Dielectrics with low loss at microwave frequencies are imperative for high-coherence solid-state quantum computing platforms. We study the dielectric loss of hexagonal boron nitride (hBN) thin films in the microwave regime by measuring the quality factor of parallel-plate capacitors (PPCs) made of NbSe$_{2}$-hBN-NbSe$_{2}$ heterostructures integrated into superconducting circuits. The extracted microwave loss tangent of hBN is bounded to be at most in the mid-10$^{-6}$ range in the low temperature, single-photon regime. We integrate hBN PPCs with aluminum Josephson junctions to realize transmon qubits with coherence times reaching 25 $\mu$s, consistent with the hBN loss tangent inferred from resonator measurements. The hBN PPC reduces the qubit feature size by approximately two-orders of magnitude compared to conventional all-aluminum coplanar transmons. Our results establish hBN as a promising dielectric for building high-coherence quantum circuits with substantially reduced footprint and, with a high energy participation that helps to reduce unwanted qubit cross-talk