MilliporeSigma
  • Home
  • Search Results
  • Morphological and functional differentiation in BE(2)-M17 human neuroblastoma cells by treatment with Trans-retinoic acid.

Morphological and functional differentiation in BE(2)-M17 human neuroblastoma cells by treatment with Trans-retinoic acid.

BMC neuroscience (2013-04-20)
Devon Andres, Brian M Keyser, John Petrali, Betty Benton, Kyle S Hubbard, Patrick M McNutt, Radharaman Ray
ABSTRACT

Immortalized neuronal cell lines can be induced to differentiate into more mature neurons by adding specific compounds or growth factors to the culture medium. This property makes neuronal cell lines attractive as in vitro cell models to study neuronal functions and neurotoxicity. The clonal human neuroblastoma BE(2)-M17 cell line is known to differentiate into a more prominent neuronal cell type by treatment with trans-retinoic acid. However, there is a lack of information on the morphological and functional aspects of these differentiated cells. We studied the effects of trans-retinoic acid treatment on (a) some differentiation marker proteins, (b) types of voltage-gated calcium (Ca2+) channels and (c) Ca2+-dependent neurotransmitter ([3H] glycine) release in cultured BE(2)-M17 cells. Cells treated with 10 μM trans-retinoic acid (RA) for 72 hrs exhibited marked changes in morphology to include neurite extensions; presence of P/Q, N and T-type voltage-gated Ca2+ channels; and expression of neuron specific enolase (NSE), synaptosomal-associated protein 25 (SNAP-25), nicotinic acetylcholine receptor α7 (nAChR-α7) and other neuronal markers. Moreover, retinoic acid treated cells had a significant increase in evoked Ca2+-dependent neurotransmitter release capacity. In toxicity studies of the toxic gas, phosgene (CG), that differentiation of M17 cells with RA was required to see the changes in intracellular free Ca2+ concentrations following exposure to CG. Taken together, retinoic acid treated cells had improved morphological features as well as neuronal characteristics and functions; thus, these retinoic acid differentiated BE(2)-M17 cells may serve as a better neuronal model to study neurobiology and/or neurotoxicity.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Potassium chloride, puriss. p.a., reag. ISO, reag. Ph. Eur., 99.5-100.5%
Supelco
Glycine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Phosgene solution, 15 wt. % in toluene
Sigma-Aldrich
Glycine, BioXtra, ≥99% (titration)
Sigma-Aldrich
Glycine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, ≥98.5%
Sigma-Aldrich
Glycine, suitable for electrophoresis, ≥99%
Sigma-Aldrich
Glycine, ReagentPlus®, ≥99% (HPLC)
Sigma-Aldrich
Calcium Ionophore A23187, ≥98% (TLC), powder
Sigma-Aldrich
Potassium chloride, BioXtra, ≥99.0%
Sigma-Aldrich
Potassium chloride solution, 0.075 M, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
Potassium chloride, for molecular biology, ≥99.0%
Sigma-Aldrich
Potassium chloride, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%
Sigma-Aldrich
Potassium chloride, AnhydroBeads, −10 mesh, 99.99% trace metals basis
Sigma-Aldrich
Potassium chloride, AnhydroBeads, −10 mesh, 99.999% trace metals basis
Sigma-Aldrich
Glycine, ACS reagent, ≥98.5%
Sigma-Aldrich
Potassium chloride, 99.999% trace metals basis
Sigma-Aldrich
Potassium chloride, ≥99.99% trace metals basis
Sigma-Aldrich
Glycine, 99%, FCC
Supelco
Potassium chloride solution, conductance standard B acc. to ISO 7888, 0.01 M KCl
Supelco
Potassium chloride solution, conductance standard A acc. to ISO 7888, 0.1 M KCl
Sigma-Aldrich
Potassium chloride solution, BioUltra, for molecular biology, ~1 M in H2O
Sigma-Aldrich
Glycine, BioUltra, for molecular biology, ≥99.0% (NT)
Sigma-Aldrich
Potassium chloride, tested according to Ph. Eur.
Sigma-Aldrich
Potassium chloride, BioUltra, for molecular biology, ≥99.5% (AT)
Sigma-Aldrich
Glycine, tested according to Ph. Eur.
Supelco
Potassium chloride solution, for Ag/AgCl electrodes, ~3 M KCl, saturated with silver chloride
Supelco
Potassium chloride solution, conductance standard C acc. to ISO 7888, 0.001 M KCl
Supelco
Potassium chloride solution, BioUltra, ~3 M in H2O
Supelco
ISA (ionic strength adjustment solution: 1 M KCl), 1 M KCl
Supelco
Calcium ionophore III, Selectophore, function tested