Label-Free Quantification of Cell Cycle Synchronicity of Human Neural Progenitor Cells Based on Electrophysiology Phenotypes
| Autor: | John H. Moore, Veronica Porterfield, Nathan S. Swami, Armita Salahi, Michael J. McConnell, Nadine Michel, Carlos Honrado |
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| Rok vydání: | 2020 |
| Předmět: |
Induced Pluripotent Stem Cells
Bioengineering 02 engineering and technology Biology 01 natural sciences Cell cycle phase Flow cytometry Neural Stem Cells Organoid medicine Humans Induced pluripotent stem cell Instrumentation Fluid Flow and Transfer Processes medicine.diagnostic_test Process Chemistry and Technology 010401 analytical chemistry Cell Cycle Cell cycle 021001 nanoscience & nanotechnology Neural stem cell 0104 chemical sciences Cell biology Electrophysiology Phenotype Stem cell 0210 nano-technology Cytometry |
| Zdroj: | ACS sensors. 6(1) |
| ISSN: | 2379-3694 |
| Popis: | The ability to coax human-induced pluripotent stem cells (hiPSCs) into human neural progenitor cells (hNPCs) can lead to novel drug discovery and transplant therapy platforms for neurological diseases. Since hNPCs can form organoids that mimic brain development, there is emerging interest in their label-free characterization for controlling cell composition to optimize organoid formation in three-dimensional (3D) cultures. However, this requires the ability to quantify hNPCs in heterogeneous samples with subpopulations of similar phenotype. Using high-throughput (>6000 cells per condition), single-cell impedance cytometry, we present the utilization of electrophysiology for quantification of hNPC subpopulations that are altered in cell cycle synchronicity by camptothecin (CPT) exposure. Electrophysiology phenotypes are determined from impedance magnitude and phase metrics for distinguishing each cell cycle phase, as validated by flow cytometry, for a wide range of subpopulation proportions. Using multishell dielectric models for each cell cycle phase, electrophysiology alterations with CPT dose could be predicted. This label-free detection strategy can prevent loss of cell viability to speed the optimization of cellular compositions for organoid development. |
| Databáze: | OpenAIRE |
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