Limits...
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.

Show MeSH

Related in: MedlinePlus

Cell aggregate sizes. Quantification of section area of GL15, SH-SY5Y, and cocultured (GL15 plus SH-SY5Y) cell aggregates (as indicated). Data are means ±SEM (n = 15) for the averaged areas of the aggregate sections, calculated using the ImageJ software (http://imagej.nih.gov/ij/).
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4309310&req=5

fig7: Cell aggregate sizes. Quantification of section area of GL15, SH-SY5Y, and cocultured (GL15 plus SH-SY5Y) cell aggregates (as indicated). Data are means ±SEM (n = 15) for the averaged areas of the aggregate sections, calculated using the ImageJ software (http://imagej.nih.gov/ij/).

Mentions: The SH-SY5Y cells were also cocultured with the GL15 cells in the RCCS bioreactor with the aim to reestablish a more neural-like microenvironment and thus to be closer to in vivo conditions. Initial experiments were carried out to determine if it was possible to establish viable GL15 plus SH-SY5Y cocultures in the RCCS bioreactor, henceforth referred to as GS-aggregates. GL15 and SH-SY5Y cells were thus cocultured in the RCCS bioreactor at a 1 : 1 ratio for up to 2 weeks. At the end of this period, the sizes of the GS-aggregates were similar to those of the monotypic G-aggregates and S-aggregates (Figure 7), and although the S-aggregates appeared smaller than the others, these differences did not reach significance. Cell viability assays also showed that the GS-aggregates had low levels of apoptotic and necrotic cells (data not shown).


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)

Cell aggregate sizes. Quantification of section area of GL15, SH-SY5Y, and cocultured (GL15 plus SH-SY5Y) cell aggregates (as indicated). Data are means ±SEM (n = 15) for the averaged areas of the aggregate sections, calculated using the ImageJ software (http://imagej.nih.gov/ij/).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig7: Cell aggregate sizes. Quantification of section area of GL15, SH-SY5Y, and cocultured (GL15 plus SH-SY5Y) cell aggregates (as indicated). Data are means ±SEM (n = 15) for the averaged areas of the aggregate sections, calculated using the ImageJ software (http://imagej.nih.gov/ij/).
Mentions: The SH-SY5Y cells were also cocultured with the GL15 cells in the RCCS bioreactor with the aim to reestablish a more neural-like microenvironment and thus to be closer to in vivo conditions. Initial experiments were carried out to determine if it was possible to establish viable GL15 plus SH-SY5Y cocultures in the RCCS bioreactor, henceforth referred to as GS-aggregates. GL15 and SH-SY5Y cells were thus cocultured in the RCCS bioreactor at a 1 : 1 ratio for up to 2 weeks. At the end of this period, the sizes of the GS-aggregates were similar to those of the monotypic G-aggregates and S-aggregates (Figure 7), and although the S-aggregates appeared smaller than the others, these differences did not reach significance. Cell viability assays also showed that the GS-aggregates had low levels of apoptotic and necrotic cells (data not shown).

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