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

Lookup NU author(s): Dr Rebecca Stewart, Professor Majlinda Lako

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Abstract

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 TERA2.cl.SP12-GFP) that expresses the genetic marker, green fluorescent protein (GFP). Here, we demonstrate that TERA2.cl.SP12-GFP 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 TERA2.cl.SP12-GFP 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.


Publication metadata

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

Publication type: Article

Publication status: Published

Journal: Stem Cells and Development

Year: 2004

Volume: 13

Issue: 6

Pages: 646-657

ISSN (print): 1547-3287

ISSN (electronic): 1557-8534

Publisher: Mary Ann Liebert, Inc. Publishers

URL: http://dx.doi.org/10.1089/scd.2004.13.646

DOI: 10.1089/scd.2004.13.646

PubMed id: 15684832


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