Cardiomyocytes from human pluripotent stem cells: from laboratory curiosity to industrial biomedical platform

Cardiomyocytes from human pluripotent stem cells (hPSCs-CMs) could revolutionise biomedicine. Global burden of heart failure will soon reach USD $90bn, while unexpected cardiotoxicity underlies 28% of drug withdrawals. Advances in hPSC isolation, Cas9/CRISPR genome engineering and hPSC-CM differentiation have improved patient care, progressed drugs to clinic and opened a new era in safety pharmacology. Nevertheless, predictive cardiotoxicity using hPSC-CMs contrasts from failure to almost total success. Since this likely relates to cell immaturity, efforts are underway to use biochemical and biophysical cues to improve many of the ~ 30 structural and functional properties of hPSC-CMs towards those seen in adult CMs. Other developments needed for widespread hPSC-CM utility include subtype specification, cost reduction of large scale differentiation and elimination of the phenotyping bottleneck. This review will consider these factors in the evolution of hPSC-CM technologies, as well as their integration into high content industrial platforms that assess structure, mitochondrial function, electrophysiology, calcium transients and contractility. This article is part of a Special Issue entitled: Cardiomyocyte Biology: Integration of Developmental and Environmental Cues in the Heart edited by Marcus Schaub and Hughes Abriel

Regulation of periodontal ligament cell behavior by cyclic mechanical loading and substrate nanotexture

Item does not contain fulltextBackground: Periodontal ligament (PDL) cells play an important role in regulating osseous remodeling and ligament formation. Mechanical loading and the specific cellular environment are involved in these processes, regulating cell behavior. However, most in vitro experimental setups investigate mechanical loading or substrate texture separately and thus do not fully represent the PDL microenvironment. Therefore, the authors investigated the influence of combined mechano-topographical stimuli on PDL cell morphology, proliferation, and osteogenic and ligament differentiation. Methods: Human PDL cells were subjected to nanometric substrate patterning and cyclic tensile stress for 2 days. Cell morphology was assessed by fluorescent staining. Further, DNA content and messenger RNA expression of osteogenic (Runx2, OCN) and ligament-related (scleraxis transcription factor (SCXA), ELN) genes were determined. Results: PDL cells adapted to the topography of nanometric groove patterns, aligning parallel with the texture. When subjected to mechanical stress, cells lost their initial orientation to the nanopattern. When subjected to dual stimuli, total DNA amounts were increased at 3 days of culture. Moreover, a significant synergistic effect on upregulation of Runx2 was observed in the combined group. For ligament-related markers, SCXA and elastin expression increased with mechanical loading and decreased on nanopatterned surfaces. Conclusion: These results suggest that mechanical stimulation is crucial in regulating periodontal cell behavior, through modulation of osteogenic and ligament gene activity, while extracellular matrix-resembling structures induce different responses from PDL cells in morphology and gene expression

The interaction between nanoscale surface features and mechanical loading and its effect on osteoblast-like cells behavior

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Substrate nanotexture and hypergravity through centrifugation enhance initial osteoblastogenesis

Item does not contain fulltextMimicking the structural nanomolecular extracellular matrix with synthetically designed nanosized materials is a relatively new approach, which can be applied in the field of bone tissue engineering. Likewise, bone tissue-engineered constructs can be aided in their development by the use of several types of mechanical stimuli. In this study, we wanted to combine nanotextured biomaterials and centrifugation in one multifactorial system. Mesenchymal stem cells were isolated from rat bone marrow, and cultured on a nanogrooved polystyrene substrate (200-nm-wide pitch with a depth of 50 nm). Constant centrifugation of 10 g was applied to cells up to 7 days. Results showed that on a nanogrooved substrate osteoblast-like cells align parallel to the groove direction. Centrifugation of 10 g also affected cell morphology on a smooth surface. Moreover, cell alignment was significantly reduced for cells grown on nanogrooved substrates, which were subsequently subjected to centrifugation. Independently, both stimuli increased the number of cells after 7 days of culture. However, when both stimuli were combined, an additive effect on cell number was observed, followed by an enhanced effect on osteocalcin mRNA expression and matrix mineralization. In conclusion, biomaterial surface modification as well as centrifugation are effective means to enhance bone cell behavior, moreover, readily available to many tissue engineers