Human embryonal carcinoma stem cells expressing green fluorescent protein form functioning neurons in vitro: A research tool for co-culture studies

Author(s): Stewart R, Coyne L, Lako M, Halliwell RF, Przyborski SA


Neural differentiation is controlled by complex molecular mechanisms that determine cell fate and diversity within the nervous system. Interactions between developing tissues play an important role in regulating this process. In vitro co-culture experiments offer a method to study cell differentiation and function under controlled conditions, with the additional benefit of investigating how interactions between populations of cells influence cell growth and behavior. However, it can often be difficult to distinguish between populations of co-cultured cells. Here we report the development of a human embryonal carcinoma (EC) stem cell line (named that expresses the genetic marker, green fluorescent protein (GFP). Here, we demonstrate that stem cells stably express GFP and that this remains detectable during retinoic acid-induced differentiation. Regulated expression of neural markers during cell development correlated with the formation of morphologically identifiable neurons. Populations of post-mitotic GFP-positive neurons were readily purified and electrophysiological characterization confirmed that such neurons were functionally active. Thus, cultured cells can be readily distinguished from alternative cell types in vitro and provide an amenable system for live cell imaging to study the development and function of human neurons in isolation, and in co-culture with other tissue types.

Similar Articles

Developmental neurotoxicity of industrial chemicals

Author(s): Grandjean P, Landrigan PJ

Neurobehavioural effects of developmental toxicity

Author(s): Grandjean P, Landrigan PJ

A tiered approach to systemic toxicity testing for agricultural chemical safety assessment

Author(s): Doe JE, Boobis AR, Blacker A, Dellarco V, Doerrer NG, et al.

Human in vitro reporter model of neuronal development and early differentiation processes

Author(s): Couillard-Despres S, Quehl E, Altendorfer K, Bentley K, Brady AM, et al.

A human pluripotent stem cell platform for assessing developmental neural toxicity screening

Author(s): Hou Z, Zhang J, Schwartz MP, Stewart R, Page CD, et al.

Towards in vitro DT/DNT testing: Assaying chemical susceptibility in early differentiating NT2 cells

Author(s): Menzner AK, AbolpourMofrad S, Friedrich O, Gilbert DF

Neuropharmacological properties of neurons derived from human stem cells

Author(s): Coyne L, Shan M, Przyborski SA, Hirakawa R, Halliwell RF

Optimizing neuronal differentiation of human pluripotent NT2 stem cells in monolayer cultures

Author(s): AbolpourMofrad S, Kuenzel K, Friedrich O, Gilbert DF