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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

Histochemical analysis of HepG2 cells cultured in monolayers (a,b); samples of HCC (c,d) and HepG2 cells cultured inside PVA/G hydrogels (e,f). For each sample type, H&E (a,c,e) and PAS reaction (b,d,f) are shown. Arrows in (b,d,f) indicate evidence of PAS positivity. S1, S2 and S3 in (e,f) define the areas of different morphotype localization in cell/scaffold constructs.
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jfb-06-00016-f003: Histochemical analysis of HepG2 cells cultured in monolayers (a,b); samples of HCC (c,d) and HepG2 cells cultured inside PVA/G hydrogels (e,f). For each sample type, H&E (a,c,e) and PAS reaction (b,d,f) are shown. Arrows in (b,d,f) indicate evidence of PAS positivity. S1, S2 and S3 in (e,f) define the areas of different morphotype localization in cell/scaffold constructs.

Mentions: Morphological analysis of cellular samples were performed using hematoxylin and eosin (H&E) staining (Figure 3a,c,e), while Periodic Acid Shiff (PAS) reaction was used to highlight the presence of glycoproteins and glycogen, as a product of metabolically active hepatic cells (Figure 3b,d,f). Glycoproteins are a class of adhesive proteins that own binding sites enabling cell attachment to the ECM. Specifically, histologic analyses were performed on HepG2 cells, samples of HCC and HepG2 cells cultured inside PVA/G hydrogels. In 2D cultures, HepG2 cells showed a typical stellate shape when distributed at a low density, while a round shape when assembled in cluster-like formations (Figure 3a). In both morphotypes, PAS reaction revealed a weak positivity as only a few cytoplasmatic granules were stained (Figure 3b). The PAS positivity was also observed on tumor sections, in which most cells displayed diffuse staining, while only a reduced number of cells were strongly positive (Figure 3d).


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)

Histochemical analysis of HepG2 cells cultured in monolayers (a,b); samples of HCC (c,d) and HepG2 cells cultured inside PVA/G hydrogels (e,f). For each sample type, H&E (a,c,e) and PAS reaction (b,d,f) are shown. Arrows in (b,d,f) indicate evidence of PAS positivity. S1, S2 and S3 in (e,f) define the areas of different morphotype localization in cell/scaffold constructs.
© Copyright Policy
Related In: Results  -  Collection

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

jfb-06-00016-f003: Histochemical analysis of HepG2 cells cultured in monolayers (a,b); samples of HCC (c,d) and HepG2 cells cultured inside PVA/G hydrogels (e,f). For each sample type, H&E (a,c,e) and PAS reaction (b,d,f) are shown. Arrows in (b,d,f) indicate evidence of PAS positivity. S1, S2 and S3 in (e,f) define the areas of different morphotype localization in cell/scaffold constructs.
Mentions: Morphological analysis of cellular samples were performed using hematoxylin and eosin (H&E) staining (Figure 3a,c,e), while Periodic Acid Shiff (PAS) reaction was used to highlight the presence of glycoproteins and glycogen, as a product of metabolically active hepatic cells (Figure 3b,d,f). Glycoproteins are a class of adhesive proteins that own binding sites enabling cell attachment to the ECM. Specifically, histologic analyses were performed on HepG2 cells, samples of HCC and HepG2 cells cultured inside PVA/G hydrogels. In 2D cultures, HepG2 cells showed a typical stellate shape when distributed at a low density, while a round shape when assembled in cluster-like formations (Figure 3a). In both morphotypes, PAS reaction revealed a weak positivity as only a few cytoplasmatic granules were stained (Figure 3b). The PAS positivity was also observed on tumor sections, in which most cells displayed diffuse staining, while only a reduced number of cells were strongly positive (Figure 3d).

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