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RCCS bioreactor-based modelled microgravity induces significant changes on in vitro 3D neuroglial cell cultures.

Morabito C, Steimberg N, Mazzoleni G, Guarnieri S, Fanò-Illic G, Mariggiò MA - Biomed Res Int (2015)

Bottom Line: We propose a human-derived neuro-/glial cell three-dimensional in vitro model to investigate the effects of microgravity on cell-cell interactions.Moreover, compared to cells as traditional static monolayers, cell aggregates cultured under modelled microgravity showed increased expression of specific differentiation markers (e.g., GL15 cells: GFAP, S100B; SH-SY5Y cells: GAP43) and modulation of functional cell-cell interactions (e.g., N-CAM and Cx43 expression and localisation).In conclusion, this culture model opens a wide range of specific investigations at the molecular, biochemical, and morphological levels, and it represents an important tool for in vitro studies into dynamic interactions and responses of nervous system cell components to microgravity environmental conditions.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroscience, Imaging and Clinical Sciences, Unit of Functional Biotechnology, Aging Research Center (Ce.S.I.), "G. d'Annunzio" University of Chieti-Pescara, Via dei Vestini 29, 66100 Chieti, Italy ; Interuniversity Institute of Myology, Italy.

ABSTRACT
We propose a human-derived neuro-/glial cell three-dimensional in vitro model to investigate the effects of microgravity on cell-cell interactions. A rotary cell-culture system (RCCS) bioreactor was used to generate a modelled microgravity environment, and morphofunctional features of glial-like GL15 and neuronal-like SH-SY5Y cells in three-dimensional individual cultures (monotypic aggregates) and cocultures (heterotypic aggregates) were analysed. Cell survival was maintained within all cell aggregates over 2 weeks of culture. Moreover, compared to cells as traditional static monolayers, cell aggregates cultured under modelled microgravity showed increased expression of specific differentiation markers (e.g., GL15 cells: GFAP, S100B; SH-SY5Y cells: GAP43) and modulation of functional cell-cell interactions (e.g., N-CAM and Cx43 expression and localisation). In conclusion, this culture model opens a wide range of specific investigations at the molecular, biochemical, and morphological levels, and it represents an important tool for in vitro studies into dynamic interactions and responses of nervous system cell components to microgravity environmental conditions.

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Expression of neuronal cell markers. Representative Western blotting and quantification of the levels of N-CAM, GAP43, and tyrosine hydroxylase (TH) in SH-SY5Y cells cultured in 2D monolayers and in the RCCS bioreactor for 24 h, 48 h, 96 h, and 2 weeks (2 w). Data are densitometric ratio analyses as means ±SEM from 3 independent experiments. *P < 0.05 and **P < 0.01 versus 2D monolayers.
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fig6: Expression of neuronal cell markers. Representative Western blotting and quantification of the levels of N-CAM, GAP43, and tyrosine hydroxylase (TH) in SH-SY5Y cells cultured in 2D monolayers and in the RCCS bioreactor for 24 h, 48 h, 96 h, and 2 weeks (2 w). Data are densitometric ratio analyses as means ±SEM from 3 independent experiments. *P < 0.05 and **P < 0.01 versus 2D monolayers.

Mentions: Western blotting carried out for the S-aggregates showed that N-CAM-140 and GAP43 expression levels were increased during the incubation, compared to the SH-SY5Y cells as 2D static monolayer cultures (Figure 6). In particular, for the S-aggregates, N-CAM-140 reached a peak after 2 weeks, while GAP43 peaked after 48 h. Conversely, N-CAM-180 and tyrosine hydroxylase did not significantly change in the S-aggregates compared to SH-SY5Y cells as 2D static cultures (Figure 6).


RCCS bioreactor-based modelled microgravity induces significant changes on in vitro 3D neuroglial cell cultures.

Morabito C, Steimberg N, Mazzoleni G, Guarnieri S, Fanò-Illic G, Mariggiò MA - Biomed Res Int (2015)

Expression of neuronal cell markers. Representative Western blotting and quantification of the levels of N-CAM, GAP43, and tyrosine hydroxylase (TH) in SH-SY5Y cells cultured in 2D monolayers and in the RCCS bioreactor for 24 h, 48 h, 96 h, and 2 weeks (2 w). Data are densitometric ratio analyses as means ±SEM from 3 independent experiments. *P < 0.05 and **P < 0.01 versus 2D monolayers.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4309310&req=5

fig6: Expression of neuronal cell markers. Representative Western blotting and quantification of the levels of N-CAM, GAP43, and tyrosine hydroxylase (TH) in SH-SY5Y cells cultured in 2D monolayers and in the RCCS bioreactor for 24 h, 48 h, 96 h, and 2 weeks (2 w). Data are densitometric ratio analyses as means ±SEM from 3 independent experiments. *P < 0.05 and **P < 0.01 versus 2D monolayers.
Mentions: Western blotting carried out for the S-aggregates showed that N-CAM-140 and GAP43 expression levels were increased during the incubation, compared to the SH-SY5Y cells as 2D static monolayer cultures (Figure 6). In particular, for the S-aggregates, N-CAM-140 reached a peak after 2 weeks, while GAP43 peaked after 48 h. Conversely, N-CAM-180 and tyrosine hydroxylase did not significantly change in the S-aggregates compared to SH-SY5Y cells as 2D static cultures (Figure 6).

Bottom Line: We propose a human-derived neuro-/glial cell three-dimensional in vitro model to investigate the effects of microgravity on cell-cell interactions.Moreover, compared to cells as traditional static monolayers, cell aggregates cultured under modelled microgravity showed increased expression of specific differentiation markers (e.g., GL15 cells: GFAP, S100B; SH-SY5Y cells: GAP43) and modulation of functional cell-cell interactions (e.g., N-CAM and Cx43 expression and localisation).In conclusion, this culture model opens a wide range of specific investigations at the molecular, biochemical, and morphological levels, and it represents an important tool for in vitro studies into dynamic interactions and responses of nervous system cell components to microgravity environmental conditions.

View Article: PubMed Central - PubMed

Affiliation: Department of Neuroscience, Imaging and Clinical Sciences, Unit of Functional Biotechnology, Aging Research Center (Ce.S.I.), "G. d'Annunzio" University of Chieti-Pescara, Via dei Vestini 29, 66100 Chieti, Italy ; Interuniversity Institute of Myology, Italy.

ABSTRACT
We propose a human-derived neuro-/glial cell three-dimensional in vitro model to investigate the effects of microgravity on cell-cell interactions. A rotary cell-culture system (RCCS) bioreactor was used to generate a modelled microgravity environment, and morphofunctional features of glial-like GL15 and neuronal-like SH-SY5Y cells in three-dimensional individual cultures (monotypic aggregates) and cocultures (heterotypic aggregates) were analysed. Cell survival was maintained within all cell aggregates over 2 weeks of culture. Moreover, compared to cells as traditional static monolayers, cell aggregates cultured under modelled microgravity showed increased expression of specific differentiation markers (e.g., GL15 cells: GFAP, S100B; SH-SY5Y cells: GAP43) and modulation of functional cell-cell interactions (e.g., N-CAM and Cx43 expression and localisation). In conclusion, this culture model opens a wide range of specific investigations at the molecular, biochemical, and morphological levels, and it represents an important tool for in vitro studies into dynamic interactions and responses of nervous system cell components to microgravity environmental conditions.

Show MeSH
Related in: MedlinePlus