Human embryonal carcinoma stem cells expressing green fluorescent protein form functioning neurons in vitro: a research tool for co-culture studies.
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.
Stewart
R
R
School of Biological and Biomedical Science, University of Durham, South Road, Durham DH1 3LE, UK.
Coyne
L
L
Lako
M
M
Halliwell
R F
RF
Przyborski
S A
SA
D018236:Carcinoma, Embryonal / Q000378:metabolism*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000378:metabolism; D005998:Glycine / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D018408:Patch-Clamp Techniques; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors; D014212:Tretinoin / Q000378:metabolism
D049452:Green Fluorescent Proteins; D014212:Tretinoin; D005998:Glycine