Autofluorescence imaging for human pluripotent stem cell differentiation into cardiomyocytes

AbstractHuman pluripotent stem cell (hPSC)-derived cardiomyocytes provide a promising regenerative cell therapy for cardiovascular patients and an important model system to accelerate drug discovery. However, cost-effective and time-efficient platforms must be developed to evaluate the quality of hPSC-derived cardiomyocytes during biomanufacturing. Here, we develop a non-invasive label-free live cell imaging platform to predict the efficiency of hPSC differentiation into cardiomyocytes. Autofluorescence imaging of metabolic co-enzymes is performed under varying differentiation conditions (cell density, concentration of Wnt signaling activator) across five hPSC lines. Live cell autofluorescence imaging and multivariate classification models provide high accuracy to separate low (< 50%) and high (≥ 50%) differentiation efficiency groups (quantified by cTnT expression on day 12) within 1 day after initiating differentiation (area under the receiver operating characteristic curve, 0.91). This non-invasive and label-free method could be used to avoid batch-to-batch and line-to-line variability in cell manufacturing from hPSCs.

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Label-free imaging for quality control of cardiomyocyte differentiation

10.1101/2021.06.01.446615 ◽

2021 ◽

Author(s):

Tongcheng Qian ◽

Tiffany Heaster ◽

Angela Houghtaling ◽

Kexin Sun ◽

Kayvan Samimi ◽

...

Keyword(s):

Characteristic Curve ◽

Cost Effective ◽

Live Cell ◽

Autofluorescence Imaging ◽

Label Free ◽

Cardiomyocyte Differentiation ◽

Non Invasive ◽

Cardiovascular Patients ◽

Density Concentration

Human pluripotent stem cell (hPSC)-derived cardiomyocytes provide a promising regenerative cell therapy for cardiovascular patients and an important model system to accelerate drug discovery. However, cost-effective and time-efficient platforms must be developed to evaluate the quality of hPSC-derived cardiomyocytes during biomanufacturing. Here, we developed a non-invasive label-free live cell imaging platform to predict the efficiency of hPSC differentiation into cardiomyocytes. Autofluorescence imaging of metabolic co-enzymes was performed under varying differentiation conditions (cell density, concentration of Wnt signaling activator) across three hPSC lines. Live cell autofluorescence imaging and multivariate classification models provided high accuracy to separate low (< 50%) and high (≥ 50%) differentiation efficiency groups (quantified by cTnT expression on day 12) within 1 day after initiating differentiation (area under the receiver operating characteristic curve, 0.98). This non-invasive and label-free method could be used to avoid batch-to-batch and line-to-line variability in cell manufacturing from hPSCs.

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Label-free imaging for cardiomyocyte differentiation from human pluripotent stem cells

10.21203/rs.3.pex-1571/v1 ◽

2021 ◽

Author(s):

Tongcheng Qian ◽

Amani Gillette ◽

Melissa Skala

Keyword(s):

Pluripotent Stem Cells ◽

Live Cell Imaging ◽

Cost Effective ◽

Autofluorescence Imaging ◽

Label Free ◽

Cardiomyocyte Differentiation ◽

Non Invasive ◽

Cardiovascular Patients ◽

Density Concentration

Abstract Human pluripotent stem cell (hPSC)-derived cardiomyocytes provide a promising regenerative cell therapy for cardiovascular patients and an important model system to accelerate drug discovery. However, cost-effective and time-efficient platforms must be developed to evaluate the quality of hPSC-derived cardiomyocytes during biomanufacturing. Here, we develop a non-invasive label-free live cell imaging platform to monitor hPSC differentiation into cardiomyocytes. Autofluorescence imaging of metabolic co-enzymes is performed under varying differentiation conditions (cell density, concentration of Wnt signaling activator) across five hPSC lines.

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