Cell Health & Viability Assays

Assessing cell viability is an essential component of any complete functional modulation or toxicity study. StemoniX offers multiple, distinct viability assays in biologically-relevant human neural-based cultures to best fit specific assessment needs. 

CellTiter-GLO® provides accurate and rapid assessment of cell viability as a function of intracellular ATP levels using a luciferin-based substrate. Cell lysis is required for this assay, so only a single viability readout can be extracted from each well.

Lactate Dehydrogenase-GLO (LDH-GLO™) quantifies cell death as the amount of LDH that has been released into the culture media upon individual cell death and lysis. Because cell lysis is not required for this assay, LDH-GLO enables longitudinal viability readouts over multiple days/treatments.

 

Image-based viability assessment provides multiparametric data surrounding culture health and morphology. First, a nuclear marker is used to label all cells present in the culture, enabling quantitative image analysis and nuclear morphological measurements that could be indicative of pyknosis or other apoptotic processes. Next, Calcein AM signal provides a direct measurement of viability when it is converted to a fluorescent derivative that gets trapped inside living cells. Finally, ethidium homodimer-1 labels only dead or dying cells with compromised plasma membrane integrity. Similar to the CellTiter-GLO assay, this image-based viability assay is a terminal assay and is only available at a single time point per culture well. However, multiple parameters describing signal intensities, nuclear morphology, cell numbers, as well as % live and % dead cells can be extracted from a single experiment.

Our  DaaS  team will work with you to determine the most suitable viability assays for your needs and identify potential multiplexed functional or toxicity assays to provide the most comprehensive answers for actionable data.  

 

Toxicology

Morphological & Structural
     Toxicity

Cardiotoxicity

Structural & 
     Gene Expression

Induced Disease

Patient-derived

Genetic engineered

AAV-based

CellTiter-GLO

General Procedure:

  1. microBrain assay platform in 96 or 384-well format.

  2. Cultures are treated with test compounds for indicated time periods. 

  3. Following compound incubation, cells are lysed using the Promega CellTiter-Glo one-step reagent.  

  4. Data are exported, normalized to vehicle control, and analyzed to provide dose response curves and IC50/EC50 values for each test article. 

 
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Oncology Disease Models

Glioblastoma

Screening

Custom Cell Lines

Expansion & Banking

Custom Growth & Assays

Assay Components

single figure with three curves; solvent control, test article, and positive control

Need data collected from the same sample over multiple timepoints? Check out our cell viability assay using LDH-Glo.

LDH-GLO

General Procedure:

  1. microBrain assay platform in 96 or 384-well format.

  2. Cultures are treated with test compounds for indicated time periods.

  3. Cell supernatant is collected from each well at the desired timepoints during or after test article treatment.

  4. LDH standard curve is prepared, and assay is run using the Promega LDH-Glo kit.

  5. Luminescence data are exported, converted to an absolute concentration of LDH (mU/mL), and analyzed to provide dose response curves and EC50/IC50s for each tested compound as appropriate.

 
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Assay Components
Image

Prefer a high content screening approach to viability? Check out our image-based viability assay.

 

Image-based Viability Studies

General Procedure:

  1. microBrain assay platform in 96 or 384-well plate format.

  2. Cultures are treated with test compounds for indicated time periods.

  3. Cells are labeled using a combination of

    • Nuclear marker – Hoechst 33342 (blue)

    • Live cell cytosolic marker - Calcein AM (green)

    • Dead cell nuclear marker - Ethidium homodimer-1 (red)

  4. High-content imaging is performed using the Perkin Elmer Operetta CLS

  5. Data are analyzed to provide % live cells, average Calcein AM intensity by condition, and average nuclear stain intensity

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Assay Components

*Significant changes can be observed in nuclear intensity following test article treatment. This can indicate signs of nuclear condensation, apoptosis or pyknosis.

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