microheart: A screening-ready, physiologically relevant human ipsc-derived cardiomyocyte platform.
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1 : A screening-ready, physiologically relevant human ipsc-derived cardiomyocyte platform.
2 Human induced Pluripotent Stem Cells (hipscs) enable assays with human, patient and organ/system-specific cells SkinPunch Biopsy Blood Sample Genetic Re-programming Validation Disease Modelling Donor Skin Fibroblasts / PBMCs hipsc Directed Differentiation hipsc Culture Expansion Tissues/Cells for Transplantation Drug screening and safety Cardiomyocytes Neurons Smooth Muscle Hepatocytes
3 Challenges in using hipsc-derived cells in more Cost of mass production and human cell screening plates Variability of batches Lack of specific features/phenotypes Maturity? Assay sensitivity, robustness, reproducibility complex assays Comley, Drug Disc. World 2016
4 Increasing Physiological Relevance of hipsc-derived models Assay Throughput 2D Cell-based models StemoniX (2D+) 3D Organoids Bioengineered and/or Bioprinted Tissue Organs on Chips Animal Models Biological Complexity / Physiological Relevance
5 hipsc-cm geometry in regular tissue cultureware is a current limitation of the model Heart Tissue Status Quo: Unstructured Boyer et al, PLOS one, 2010 Human ips-derived Cardiomyocytes on traditional cell cultureware
6 Cutting-edge stem cell biology + Nano/micro manufacturing expertise = StemoniX Cell-Ready: Micro-engineered screening plates that allow CMs to passively self-align into structures resemblant of cardiac muscle fibers Assay-Ready: Ready-to-use differentiated hipsc-cms pre-plated for high-throughput screening in drug discovery and precision medicine
7 StemoniX promotes hipsc-cm alignment Standard cell cultureware StemoniX 10X, BF Intensity Standard cell cultureware StemoniX 40X, SaAct Light scattering angles
8 StemoniX alignment improves unidirectional contraction Standard Tissue cultureware StemoniX Cardiomyocyte Motion Tracking Huebsch et al, Nat Sci Rep, 2016
9 4X StemoniX promotes passive self-alignment in hipsc-cms Standard cell cultureware StemoniX alignment technology DNA/cTNT/Sarcomeric aact 60X
10 StemoniX enhances sarcomeric alignment in hipsc-cms Standard cell cultureware StemoniX alignment technology DNA/cTNT/Sarcomeric aact
11 StemoniX promotes targeting of cardiac cell junction components to distal intercalated discs Standard Tissue cultureware DNA / Plakoglobin StemoniX
12 StemoniX induces an increase in the expression levels of key cardiac physiology genes Relative expression Standard Cell Cultureware StemoniX RYR2 ATP2A2 PLN CACNA1C SCN5A KCNE1 KCNQ1 GJA1 GJA5 DSP TBX5 MEF2C ISL1 TNNT2 NPPB Ca 2+ handling Ion channels Cell junctions Cardiac TFs
13 Time (Seconds) Time (Seconds) StemoniX is compatible with high throughput calcium flux assays Standard tissue cultureware StemoniX alignment technology Molecular Devices FLIPR Tetra Relative Light Units Relative Light Units
14 High Throughput Calcium Flux Analysis indicates more efficient calcium PSSD PRT PSSD handling of hipsc-cms in StemoniX Beat Rate () A.U Peak Amplitude Peak Amplitude (A.U.) * 2.5 Legend PSSD Peak Spacing Standard Deviation (s) A.U PW@10% Peak Rise Peak Baseline * * CV 24.99% 12.01% 18.11% 17.18% 15.63% 14.65% CV 34.87% 24.17% PDT Peak Decay Peak Spacing 1.0 Peak PW@10% Width at 10% Amplitude (s) PW@10% * *p<01
15 StemoniX is compatible with membrane potential recordings Standard Tissue Cultureware StemoniX Alignment Technology
16 StemoniX is compatible with bright light image-based label-free contraction analysis Video ROI Measurement Molecular Devices ImageXpress Micro Confocal
17 StemoniX Cellular Geometry Resembling Human Heart Tissue Heart Tissue Status Quo: Unstructured StemoniX Boyer et al, PLOS one, 2010 Human ips-derived Cardiomyocytes on traditional cell cultureware ips-cardiomyocytes on StemoniX
18 Summary StemoniX High Throughput Screening platform that more closely resembles the tissue architecture of native human heart tissue Cardiomyocyte alignment, coherent directionality of contraction More physiological cell geometry Enhanced sarcomeric organization, perpendicular alignment, intercalated disc localization Improved expression profiles of key cardiac physiology genes, including cardiomyocyte calcium handling pathways, ion channels, cardiac transcription factors and contractile proteins. More efficient calcium handling Significance offers a cardiomyocyte platform with greater physiological relevance that is pre-formatted as a high throughput screening platform A.U PSSD 0.3 Peak Amplitude (A.U.) PW@10% Peak Rise * * PSSD PW@10% Peak Width at 10% Amplitude (s) * PRT PDT Peak Decay * Legend
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