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Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study.

Moscato S, Ronca F, Campani D, Danti S - J Funct Biomater (2015)

Bottom Line: Morphological features of PVA/G hydrogels were investigated, resulting to mimic the trabecular structure of liver parenchyma.Furthermore, β-actin and α5β1 integrin revealed a morphotype-related expression; in particular, the frontline cells were characterized by a strong immunopositivity on a side border of their membrane, thus suggesting the formation of lamellipodia-like structures apt for migration.Based on these results, we propose PVA/G hydrogels as valuable substrates to develop a long term 3D HCC model that can be used to investigate important aspects of tumor biology related to migration phenomena.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Experimental Medicine, University of Pisa, via Savi 10, 56126 Pisa, Italy. stefania.moscato@unipi.it.

ABSTRACT
It has been demonstrated that three-dimensional (3D) cell culture models represent fundamental tools for the comprehension of cellular phenomena both for normal and cancerous tissues. Indeed, the microenvironment affects the cellular behavior as well as the response to drugs. In this study, we performed a morphological analysis on a hepatocarcinoma cell line, HepG2, grown for 24 days inside a bioartificial hydrogel composed of poly(vinyl alcohol) (PVA) and gelatin (G) to model a hepatocellular carcinoma (HCC) in 3D. Morphological features of PVA/G hydrogels were investigated, resulting to mimic the trabecular structure of liver parenchyma. A histologic analysis comparing the 3D models with HepG2 cell monolayers and tumor specimens was performed. In the 3D setting, HepG2 cells were viable and formed large cellular aggregates showing different morphotypes with zonal distribution. Furthermore, β-actin and α5β1 integrin revealed a morphotype-related expression; in particular, the frontline cells were characterized by a strong immunopositivity on a side border of their membrane, thus suggesting the formation of lamellipodia-like structures apt for migration. Based on these results, we propose PVA/G hydrogels as valuable substrates to develop a long term 3D HCC model that can be used to investigate important aspects of tumor biology related to migration phenomena.

No MeSH data available.


Related in: MedlinePlus

Western blot results of β-actin and HPRT expression in HepG2 cells cultured in monolayers for 4 days (80% confluence) and inside PVA/G hydrogels for 24 days. Hydrogels without cells were tested as negative controls. The immunoblot reaction is shown in (a) and the band analysis in (b).
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jfb-06-00016-f006: Western blot results of β-actin and HPRT expression in HepG2 cells cultured in monolayers for 4 days (80% confluence) and inside PVA/G hydrogels for 24 days. Hydrogels without cells were tested as negative controls. The immunoblot reaction is shown in (a) and the band analysis in (b).

Mentions: Immunoblot assay was performed on HepG2 cells cultured both in monolayers (2D) and inside PVA/G hydrogels (Figure 6). Results showed a high expression of β-actin in cell/scaffold constructs, thus confirming the previous immunohistochemical data. Moreover, the volume intensity evaluation of the bands revealed that in the cell/scaffold constructs β-actin was expressed at a higher concentration than in HepG2 monolayer cultures, taking in account that the same total protein amount was loaded and separated by the SDS-PAGE gels. Its calculated values for band volume intensity of cell/scaffold constructs and monolayer cells were 41.10 × 105 and 32.06 × 105, respectively (Figure 6b).


Poly(vinyl alcohol)/gelatin Hydrogels Cultured with HepG2 Cells as a 3D Model of Hepatocellular Carcinoma: A Morphological Study.

Moscato S, Ronca F, Campani D, Danti S - J Funct Biomater (2015)

Western blot results of β-actin and HPRT expression in HepG2 cells cultured in monolayers for 4 days (80% confluence) and inside PVA/G hydrogels for 24 days. Hydrogels without cells were tested as negative controls. The immunoblot reaction is shown in (a) and the band analysis in (b).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4384098&req=5

jfb-06-00016-f006: Western blot results of β-actin and HPRT expression in HepG2 cells cultured in monolayers for 4 days (80% confluence) and inside PVA/G hydrogels for 24 days. Hydrogels without cells were tested as negative controls. The immunoblot reaction is shown in (a) and the band analysis in (b).
Mentions: Immunoblot assay was performed on HepG2 cells cultured both in monolayers (2D) and inside PVA/G hydrogels (Figure 6). Results showed a high expression of β-actin in cell/scaffold constructs, thus confirming the previous immunohistochemical data. Moreover, the volume intensity evaluation of the bands revealed that in the cell/scaffold constructs β-actin was expressed at a higher concentration than in HepG2 monolayer cultures, taking in account that the same total protein amount was loaded and separated by the SDS-PAGE gels. Its calculated values for band volume intensity of cell/scaffold constructs and monolayer cells were 41.10 × 105 and 32.06 × 105, respectively (Figure 6b).

Bottom Line: Morphological features of PVA/G hydrogels were investigated, resulting to mimic the trabecular structure of liver parenchyma.Furthermore, β-actin and α5β1 integrin revealed a morphotype-related expression; in particular, the frontline cells were characterized by a strong immunopositivity on a side border of their membrane, thus suggesting the formation of lamellipodia-like structures apt for migration.Based on these results, we propose PVA/G hydrogels as valuable substrates to develop a long term 3D HCC model that can be used to investigate important aspects of tumor biology related to migration phenomena.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Experimental Medicine, University of Pisa, via Savi 10, 56126 Pisa, Italy. stefania.moscato@unipi.it.

ABSTRACT
It has been demonstrated that three-dimensional (3D) cell culture models represent fundamental tools for the comprehension of cellular phenomena both for normal and cancerous tissues. Indeed, the microenvironment affects the cellular behavior as well as the response to drugs. In this study, we performed a morphological analysis on a hepatocarcinoma cell line, HepG2, grown for 24 days inside a bioartificial hydrogel composed of poly(vinyl alcohol) (PVA) and gelatin (G) to model a hepatocellular carcinoma (HCC) in 3D. Morphological features of PVA/G hydrogels were investigated, resulting to mimic the trabecular structure of liver parenchyma. A histologic analysis comparing the 3D models with HepG2 cell monolayers and tumor specimens was performed. In the 3D setting, HepG2 cells were viable and formed large cellular aggregates showing different morphotypes with zonal distribution. Furthermore, β-actin and α5β1 integrin revealed a morphotype-related expression; in particular, the frontline cells were characterized by a strong immunopositivity on a side border of their membrane, thus suggesting the formation of lamellipodia-like structures apt for migration. Based on these results, we propose PVA/G hydrogels as valuable substrates to develop a long term 3D HCC model that can be used to investigate important aspects of tumor biology related to migration phenomena.

No MeSH data available.


Related in: MedlinePlus