|Cell line/type||hPSCs derived cardiomyocytes|
|Product||CDI cardiac 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 induced pluripotent stem cell (iPSc) differentiated Cardiomyocytes (Cellular Dynamics International, CDI) were plated according to user instructions and one cryovial was used for each experiment. Cardiomyocytes were plated with CDI cardiac plating medium, 7 days before transferring to the in vitro system, on control SiO2 surfaces (500 cells/mm2), or cantilevers (2,500 cells/mm2) previously coated with fibronectin (50 μ g/mL in 1X PBS (phosphate buffer solution)). Forty-eight hours later, the medium was replaced with cardiac serum-free maintenance medium, and followed by half medium changes every 48 hours before 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