Engineered Maturation Approaches of Human Pluripotent Stem Cell-Derived Ventricular Cardiomyocytes
Abstract
:1. Introduction
2. Characteristics of Mature and Immature Cardiomyocytes
3. Approaches to Obtaining Mature Cardiomyocytes
3.1. Biophysical and Biochemical Factors
3.2. Electrophysiological Stimulation
3.3. Mechanical Stress
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Mature CMs | Immature CMs | ||
---|---|---|---|
Structure | Structure | Rod-shaped | Round and irregular |
Alignment | Orderly | Disorderly | |
Nucleation | 20–30% binuclear or polynuclear | Slightly binuclear | |
Beating | Quiescent | Spontaneous | |
Length–width ratio | 5–10:1 | 1–3:1 | |
Sarcomere banding | Z discs, I band, H band, A band, M band | Z discs, I band | |
Sarcomere length | 2.2 μm | 1.6 μm | |
Troponin | cTnT, high β-MHC/α-MHC, high MLC2v/MLC2a, high cTnI/fetal ssTnI, Titin isoform N2B, ADRA1A | cTnT, low β-MHC/α-MHC, nondeterministic MLC2v/MLC2a, low cTnI/fetal ssTnI, Titin isoform N2BA | |
SRP | High CSQ, PLN, RYR2, SERCA/ATP2A2 | Low CSQ, PLN, RYR2, SERCA/ATP2A2 | |
T-tubules | Present | Not present | |
Mitochondria | Regularly distributed; 20–40% of cell volume | Irregularly distributed; paucity | |
LGJ | Intercalated discs | Circumferential | |
Biochemistry | Metabolism | Fatty acid β-oxidative | Glycolysis and lactate |
Biophysical | Force | 40–80 mN/mm2 for muscle lines μN range for a single cell | 0.08-4 mN/mm2 for 3D cultivation nN range for a single cell |
Electrophysiology | Capacitance | 150 pF | 10–30 pF |
RMP | −80 to −90 mV | −20 to −60 mV | |
Upstroke velocity | 100–300 V/s | 10–50 V/s | |
Conduction velocity | 60 cm/s | 10–20 cm/s | |
APA | 100–110 mV | 70–120 mV |
Stimuluses | Cultured Cell Types | Maturation Conditions | Reference |
---|---|---|---|
Electric stimulation | Hes3 hESCs | After 4 days of culturing in the presence of electric field stimulation (a 6.6 V/cm, 1 Hz, and 2 ms pulse), hESC-CM elongation and troponin-T enhancement. | [48] |
Hes2 and Hes3 hESCs and CDI-MRB HR-I-2Cr-2R hiPSCs | Biowires increased myofibril ultrastructural organization, elevated conduction velocity, improved Calcium handling properties, and produced better electrophysiological performance. | [42] | |
C25 hiPSCs | 2 Hz in the first week and 1.5 Hz thereafter, developed 1.5-fold contractile forces. | [49] | |
hiPSC-CMs (ReproCardio 2) | Efficient electrical stimulations were formed by a hydrogel-based microchamber with organic electrodes. The large interfacial capacitance of the electrodes eliminated cytotoxic bubbles. | [50] | |
Electric stimulation and mechanical strain | Neonatal Rat Heart Cells | Engineered heart muscle was subjected to electric stimulation at 0, 2, 4, or 6 Hz for 5 days and engineered flexible poles facilitated auxotonic contractions by straining. Force–frequency relationships of 2 and 4 Hz stimulation were divergent. | [51] |
C2A, WTC-11, IMR90, and BS2 hiPSCs | After the first contraction was observed, tissue was immediately subjected to 21 days of increasingly intense electromechanical strain. Cell properties were then evaluated by a multiplex test. | [52] | |
Mechanical loading | HES2 hESCs | Cyclical stretching produced Cardiac troponin T elevation, cell elongation, and an increase in gap junction. | [29] |
IMR90 ESCs and IBJ hiPSCs | Compared to a 2D culture, a 3D environment increased the number of cardiomyocytes and decreased the number of smooth muscles. With cyclic stress, expression of several cardiac markers increased, including β-myosin heavy chains and cardiac troponin T. | [35] | |
Mechanical loading and vascular co-culture | H7 hESCs | Cyclic stress enhanced cardiomyocyte hypertrophy and proliferation rates significantly and endothelial cells showed the formation of vessel-like structures. | [31] |
Textile based-culturing | UTA.04602 hiPSC | Gelatin-coated polyethylene terephthalate-based textiles were used as the culturing surface. hiPSC-CMs showed improved structural properties. | [53] |
Substrate stiffness | Neonatal Rat Ventricular Myocytes | Substrates of varying elastic moduli were fabricated. Cardiomyocytes matured on 10 kPa gels were similar to the native myocardium and generated a greater mechanical force and the largest calcium transients. | [28] |
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Ge, F.; Wang, Z.; Xi, J.J. Engineered Maturation Approaches of Human Pluripotent Stem Cell-Derived Ventricular Cardiomyocytes. Cells 2020, 9, 9. https://doi.org/10.3390/cells9010009
Ge F, Wang Z, Xi JJ. Engineered Maturation Approaches of Human Pluripotent Stem Cell-Derived Ventricular Cardiomyocytes. Cells. 2020; 9(1):9. https://doi.org/10.3390/cells9010009
Chicago/Turabian StyleGe, Feixiang, Zetian Wang, and Jianzhong Jeff Xi. 2020. "Engineered Maturation Approaches of Human Pluripotent Stem Cell-Derived Ventricular Cardiomyocytes" Cells 9, no. 1: 9. https://doi.org/10.3390/cells9010009