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Comparison of Human Induced Pluripotent Stem Cell-Derived Neurons and Rat Primary CorticalNeurons as In Vitro Models of Neurite Outgrowth
Druwe, I., T. Freudenrich, K. Wallace, Tim Shafer, AND W. Mundy. Comparison of Human Induced Pluripotent Stem Cell-Derived Neurons and Rat Primary CorticalNeurons as In Vitro Models of Neurite Outgrowth. Applied In Vitro Toxicology. Mary Ann Liebert, Inc., Larchmont, NY, 2(1):26-36, (2016).
In vitro assays using neural cell cultures are being developed for the key events of neurodevelopment including neurite outgrowth. While neuronal cell lines and primary rodent cultures have been widely employed, they may not fully recapitulate human biology. The present study evaluated the utility of human iPSC-derived neurons as a model for in vitro evaluation of chemicals for effects on neurite outgrowth. The results show that human iPSC-derived neurons are suitable for use in high-throughput assays of neurite outgrowth, and that the response to chemicals was different from the response observed in rat cortical neurons. This data supports the concept that in vitro assays for chemical safety testing should focus on cells of human origin.
High-throughput assays that can quantify chemical-induced changes at the cellular and molecular level have been recommended for use in chemical safety assessment. High-throughput, high content imaging assays for the key cellular events of neurodevelopment have been proposed to rapidly evaluate chemicals for developmental neurotoxicity. Thus, in vitro assays using neural cell cultures are being developed for key neurodevelopmental events including proliferation, neural differentiation, migration, synaptogenesis and neurite outgrowth. While neuronal cell lines and primary rodent cultures have been widely employed, they may not fully recapitulate human biology. The advent of human induced pluripotent stem cell (iPSC)-derived neurons may help to overcome this problem. Human iPSC-derived neurons demonstrate the morphology and physiology of their in situ counterparts and have been commercialized and made available in large numbers, facilitating their use in chemical testing. The present study evaluated the neuronal characteristics of commercially available, cryopreserved human iPSC-derived neurons (iCell® neurons) in a 96-well format. Characterization of the cells using immunocytochemistry revealed a relatively pure population of neurons that extended both putative axons and dendrites. The cells developed neurites rapidly and consistently as quantified using automated high content imaging. Exposure to a training set of chemicals previously demonstrated to affect neurite outgrowth in rodent and/or human neurons resulted in concentration-dependent decreases in neurite outgrowth in human iCell® neurons, although the response to chemicals was different from the response observed in rat cortical neurons. Lead had no effect in rat cortical neurons but decreased neurite outgrowth in human neurons. I contrast, etoposide decreased both neurite outgrowth and viability in rat primary cortical cultures, but was etoposide was much less potent in human iPSC-derived neurons. Together, these data demonstrate that commercially available human iPSC-derived neurons are amenable for use in medium- to high-throughput assays of neurite outgrowth using high content imaging. Though limited, the data indicate that human and rodent neurons can respond differently to chemical insult, and support the concept that in vitro assays for chemical safety testing focus on cells of human origin.Key words: chemical safety assessment, induced pluripotent stem cell, neurite outgrowth, high content imaging
Record Details:Record Type: DOCUMENT (JOURNAL/PEER REVIEWED JOURNAL)
Organization:U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
NATIONAL HEALTH AND ENVIRONMENTAL EFFECTS RESEARCH LABORATORY
INTEGRATED SYSTEMS TOXICOLOGY DIVISION
SYSTEMS BIOLOGY BRANCH