Turning teratocarcinoma cells into neurons: rapid differentiation of NT-2 cells in floating spheres.
Cells from the human teratocarcinoma line NTera-2 can be induced to terminally differentiate into postmitotic neurons when treated with retinoic acid. However, this differentiation process is rather time consuming as it takes between 42 and 54 days. Here, we propose a modified differentiation protocol which reduces the time needed for differentiation considerably without compromising the quantity of the neurons obtained. The introduction of a proliferation step as free floating cell spheres cuts the total time needed to obtain high yields of purified NT-2 neurons to about 24-28 days. The cells obtained show neuronal morphology and migrate to form ganglion-like cell conglomerates. Differentiated cells express neuronal polarity markers such as the cytoskeleton associated proteins MAP2 and Tau. Moreover, the generation of neurons in sphere cultures induced immunoreactivity to the ELAV-like neuronal RNA-binding proteins HuC/D, which have been implicated in mechanisms of nerve cell differentiation.
Paquet-Durand
François
F
Physiologisches Institut, Abteilung Zellbiologie, Tierärztliche Hochschule Hannover, Bischofsholer Damm 15, D-30559, Hannover, Germany.
Tan
Saime
S
Bicker
Gerd
G
D015415:Biomarkers; D002449:Cell Aggregation / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000235:genetics*; D002465:Cell Movement / Q000502:physiology; D016764:Cell Polarity / Q000502:physiology; D048429:Cell Size / Q000502:physiology; D051959:ELAV Proteins; D000067778:ELAV-Like Protein 3; D005724:Ganglia / Q000166:cytology / Q000254:growth & development / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism; D018874:Spheroids, Cellular / Q000166:cytology / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D018243:Teratocarcinoma / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism*; D016875:tau Proteins / Q000378:metabolism
D015415:Biomarkers; D051959:ELAV Proteins; D000067778:ELAV-Like Protein 3; C490264:ELAVL3 protein, human; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D016601:RNA-Binding Proteins; D016875:tau Proteins