Available Assays:


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In vitro toxicity:


Basal cytotoxicity: All compounds will be screened for basal cytotoxicity by assessing viability of HeLa cells after nanomaterial exposure for 24 hours. Viability will be assessed using MTT, which indicates mitochondrial activity.

Tissue specific toxicity: Users can specify tissue specific models of interest for a subset of compounds to assess toxicity in hepatocyte or macrophage physiology. It is predicted these two cell types will act as major sources of clearance and interaction.


In vivo toxicity:


A subset of compounds will be screened in animals for preliminary in vivo toxicity in collaboration with the animal core. Promising formulations will be submitted to NCL or Charles River for full toxicological evaluation with the cost to be borne by NCI in case of selection by NCL or the individual project budget in the case of Charles River.

Assays available at the current time are summarized below


Cell types:


Basal cytotoxicity:

Human cell line HeLa


Tissue specific toxicity:

Liver:

Random cocultured hepatocyte (mouse, rat or human)

Micropatterned co-culture hepatocyte (mouse, rat or human)


Macrophage:

Murine macrophage

Human U937 cell line



Assays:


Viability determined by MTT

Differentiated function, migration and chronic response of hepatocytes

TNF secretion and phagocytosis of macrophages.


Pilot Studies:


The nanotox core tested five compounds on HeLa, HepG2, rat hepatocytes and co-cultured rat hepatocytes, as part of a pilot study. The following TC50 results generated after 24 hours of compound exposure. Human cell lines were cultured for 24 hours and hepatocytes were cultured 7 days prior to addition of compound. Viability was determined via MTT, which shows mitochondrial function.


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Human liver specific toxicity has been tested using hepatocyte co-cultures. This is shown below


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Submitting Samples:


For the purposes of the Nanomaterials Toxicity Core a single nanomaterial formulation will constitute a ‘compound’ (ie a single core material with 6 coatings is 6 compounds).


Nanomaterial submissions must be provided in a water soluble format and should be stable in solution for at least one week.


Core projects will be given priority over pilot projects.


Approximate wait time will be 2-4 weeks.


To submit samples, please Log in and complete the online submission form. Please complete all required fields and provide any relevant information (clinical concentration, range you want tested, supply is limited etc).


Samples will then be tested in triplicate at various concentrations depending on the nanomaterial submitted, using a log scale, and a doubling scale. For example iron oxide particles used at a clinical concentration of 0.2mg/ml requires at least 76.8mg in 600ul for each cell type you wish to be tested if the highest concentration used is 20mg/ml (100x). If supply is limited then the range will be decreased as needed. (ie use 16.8mg in 600ul only allows for 2mg/ml (10x) as the highest concentration).

If you need any assistance please email nanotoxcore@mit.edu