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The Effect of Coatings and Nerve Growth Factor on Attachment and Differentiation of Pheochromocytoma Cells

Cellular attachment plays a vital role in the differentiation of pheochromocytoma (PC12) cells. PC12 cells are noradrenergic clonal cells isolated from the adrenal medulla of Rattus norvegicus and studied extensively as they have the ability to differentiate into sympathetic neuron-like cells. The e...

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Autores principales: Orlowska, Anna, Perera, Pallale Tharushi, Al Kobaisi, Mohammad, Dias, Andre, Nguyen, Huu Khuong Duy, Ghanaati, Shahram, Baulin, Vladimir, Crawford, Russell J., Ivanova, Elena P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5793558/
https://www.ncbi.nlm.nih.gov/pubmed/29301234
http://dx.doi.org/10.3390/ma11010060
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author Orlowska, Anna
Perera, Pallale Tharushi
Al Kobaisi, Mohammad
Dias, Andre
Nguyen, Huu Khuong Duy
Ghanaati, Shahram
Baulin, Vladimir
Crawford, Russell J.
Ivanova, Elena P.
author_facet Orlowska, Anna
Perera, Pallale Tharushi
Al Kobaisi, Mohammad
Dias, Andre
Nguyen, Huu Khuong Duy
Ghanaati, Shahram
Baulin, Vladimir
Crawford, Russell J.
Ivanova, Elena P.
author_sort Orlowska, Anna
collection PubMed
description Cellular attachment plays a vital role in the differentiation of pheochromocytoma (PC12) cells. PC12 cells are noradrenergic clonal cells isolated from the adrenal medulla of Rattus norvegicus and studied extensively as they have the ability to differentiate into sympathetic neuron-like cells. The effect of several experimental parameters including (i) the concentration of nerve growth factor (NGF); (ii) substratum coatings, such as poly-L-lysine (PLL), fibronectin (Fn), and laminin (Lam); and (iii) double coatings composed of PLL/Lam and PLL/Fn on the differentiation process of PC12 cells were studied. Cell morphology was visualised using brightfield phase contrast microscopy, cellular metabolism and proliferation were quantified using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, and the neurite outgrowth and axonal generation of the PC12 cells were evaluated using wide field fluorescence microscopy. It was found that double coatings of PLL/Lam and PLL/Fn supported robust adhesion and a two-fold enhanced neurite outgrowth of PC12 cells when treated with 100 ng/mL of NGF while exhibiting stable metabolic activity, leading to the accelerated generation of axons.
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spelling pubmed-57935582018-02-07 The Effect of Coatings and Nerve Growth Factor on Attachment and Differentiation of Pheochromocytoma Cells Orlowska, Anna Perera, Pallale Tharushi Al Kobaisi, Mohammad Dias, Andre Nguyen, Huu Khuong Duy Ghanaati, Shahram Baulin, Vladimir Crawford, Russell J. Ivanova, Elena P. Materials (Basel) Article Cellular attachment plays a vital role in the differentiation of pheochromocytoma (PC12) cells. PC12 cells are noradrenergic clonal cells isolated from the adrenal medulla of Rattus norvegicus and studied extensively as they have the ability to differentiate into sympathetic neuron-like cells. The effect of several experimental parameters including (i) the concentration of nerve growth factor (NGF); (ii) substratum coatings, such as poly-L-lysine (PLL), fibronectin (Fn), and laminin (Lam); and (iii) double coatings composed of PLL/Lam and PLL/Fn on the differentiation process of PC12 cells were studied. Cell morphology was visualised using brightfield phase contrast microscopy, cellular metabolism and proliferation were quantified using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, and the neurite outgrowth and axonal generation of the PC12 cells were evaluated using wide field fluorescence microscopy. It was found that double coatings of PLL/Lam and PLL/Fn supported robust adhesion and a two-fold enhanced neurite outgrowth of PC12 cells when treated with 100 ng/mL of NGF while exhibiting stable metabolic activity, leading to the accelerated generation of axons. MDPI 2017-12-31 /pmc/articles/PMC5793558/ /pubmed/29301234 http://dx.doi.org/10.3390/ma11010060 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Orlowska, Anna
Perera, Pallale Tharushi
Al Kobaisi, Mohammad
Dias, Andre
Nguyen, Huu Khuong Duy
Ghanaati, Shahram
Baulin, Vladimir
Crawford, Russell J.
Ivanova, Elena P.
The Effect of Coatings and Nerve Growth Factor on Attachment and Differentiation of Pheochromocytoma Cells
title The Effect of Coatings and Nerve Growth Factor on Attachment and Differentiation of Pheochromocytoma Cells
title_full The Effect of Coatings and Nerve Growth Factor on Attachment and Differentiation of Pheochromocytoma Cells
title_fullStr The Effect of Coatings and Nerve Growth Factor on Attachment and Differentiation of Pheochromocytoma Cells
title_full_unstemmed The Effect of Coatings and Nerve Growth Factor on Attachment and Differentiation of Pheochromocytoma Cells
title_short The Effect of Coatings and Nerve Growth Factor on Attachment and Differentiation of Pheochromocytoma Cells
title_sort effect of coatings and nerve growth factor on attachment and differentiation of pheochromocytoma cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5793558/
https://www.ncbi.nlm.nih.gov/pubmed/29301234
http://dx.doi.org/10.3390/ma11010060
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