|Cell line/type||iPSc differentiated cortical-like neurons|
|Product||CDI neuronal plating medium|
From: Oleaga, C., Bernabini, C., Smith, A. S., Srinivasan, B., Jackson, M., McLamb, W., ... & Berry, B. (2016). Multi-Organ toxicity demonstration in a functional human in vitro system composed of four organs. Scientific reports, 6, 20030.
In this study, a functional human model is used to evaluate multi-organ toxicity in a 4-organ system under continuous flow conditions in a serum-free defined medium. The cell types and corresponding media used in this study are described seperately in the FCS-free Database. The other cell types are:
Human iPSc differentiated cortical-like neurons (Cellular Dynamics International, CDI) were plated according to user instructions and one cryovial was used for each experiment. Neurons were plated with CDI neuronal plating medium, 2–4 days before transferring to the in vitro system, cells were plated onto DETA modified 15 mm diameter round glass coverslips and then coated with laminin (3.3 μ g/mL) at a density of 700 cells/mm2. Forty-eight hours later, the medium was replaced with cortical serum-free maintenance medium, followed by half medium changes every 48 hours before the co-culture.
The defined serum-free medium base composition used to maintain the functionality of the 4 organ culture system for up to 14 days was previously published62 and was modified by the addition of an antibiotic/antimycotic cocktail at 1X (Life Technologies, 15240-062) and the elimination of the G5 supplement. The base medium was composed of 1X Neurobasal (Life Technologies, 21103-049), 1X B27 (Life Technologies, 17504-044), 1X Antibiotic and antimycotic (Life Technologies, 15240-062) and 1X Glutamax (Life Technologies, 35050-061). It was supplemented with 10 ng/mL Glial-derived Neurotrophic Factor (GDNF) (Cell Sciences, CRG400B), 20 ng/mL Brain-derived Neurotrophic Factor (BDNF) (Cell Sciences, CRB600B), 5 ng/mL Ciliary-derived Neurotrophic Factor (CNTF) (Cell Sciences, CRC400A), 20 ng/mL Neurotrophin-3 and 4 (NT3 and NT4) (Cell Sciences, CRN500B and CRN501B), 100 ng/mL Vitronectin (Sigma, V8379-50UG), 10 ng/mL Insulin-like Growth Factor-I (IGF-1) (PeproTech, 100-11), 100 ng/mL Agrin (R&D, 550-AG-100), 1 μ M Adenosine 3′ , 5′ -cyclic monophosphate (cAMP) (Sigma, A9501), 4 μ g/mL laminin (Life Technologies 23017-015), 50 ng/mL Sonic Hedgehog, N-terminal peptide (Shh) (R&D, 1845-SH-025) and 0.1 μ M Retinoic acid (Sigma, R2625). All the compounds purchased as a powder were dissolved in water, except for retinoic acid which was dissolved in DMEM 1X.https://www.nature.com/articles/srep20030
|Source||Literature - modified commercial product|
|Chemically defined > No||No|
Oleaga et al (2016) - Oleaga, C., Bernabini, C., Smith, A. S., Srinivasan, B., Jackson, M., McLamb, W., ... & Berry, B. (2016). Multi-Organ toxicity demonstration in a functional human in vitro system composed of four organs. Scientific reports, 6, 20030