Table 1 Definitions of QC criteria for motor neuronal cultures; cells assessed at neural precursor cell stage (NPC) and after terminal differentiation for 3 days (D3) or 10 days (D10) to produce early and later motor neurons respectively; image analysis scripts run in signals image artist (Revvity).
From: Examining iPSC derived motor neuron variability and genome stability monitoring as a solution
QC Metric Class | QC Metric | QC metric description | Assessment tool | Criteria for passing |
|---|---|---|---|---|
Cell Morphology | Non neuronal morphology? | Clear overcrowding of images with non neuronal cells? | Operator survey of image | Clear neuronal images |
Clusters > 5000 μm²? | Images with cellular clusters greater than 5000 μm² non amenable to automated accurate image analysis? | Image Analysis Script | No clusters | |
Cell Proliferation | NPC: D3 | Cell number ratio of D3 (early stage motor neurons) to NPCs (neural precursors) | R script & Image Analysis Script | < 3 |
D3:D10 | Cell number ratio of D10 (late stage motor neurons) to D3 (early stage motor neurons) | R script & Image Analysis Script | < 1 | |
Neuronal Health | D10 neurite area (µm²) | Sum of neurite area per well | Image Analysis Script | > 40,000 |
Culture Purity | PAX6 + OLIG2 (%) | Co-expression of neural precursor markers | Image Analysis Script | > 80 |
SMI32 + MAP2 (%) | Co-expression of motor neuronal with pan neuronal markers | Image Analysis Script | ||
ISL1 + MAP2 (%) | Co-expression of motor neuronal with pan neuronal markers | Image Analysis Script |
