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A hybrid substratum for primary hepatocyte culture that enhances hepatic functionality with low serum dependency.

Meng Q, Tao C, Qiu Z, Akaike T, Cui F, Wang X - Int J Nanomedicine (2015)

Bottom Line: A poly-(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide) matrix can recognize cells and promote liver function in a spheroid structure because of a specific galactose-asialoglycoprotein receptor interaction.Meanwhile, a fusion protein, E-cadherin-Fc, when incubated with various cells, has shown an enhancing effect on cellular viability and metabolism.The isolated cells showed a monolayer aggregate morphology on the coimmobilized surface and displayed higher functional expression than cells on traditional matrices.

View Article: PubMed Central - PubMed

Affiliation: School of Materials Science and Engineering, Tsinghua University, Beijing, People's Republic of China ; State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, People's Republic of China ; Biomaterials Center for Regenerative Medical Engineering, Ibaraki, Japan.

ABSTRACT
Cell culture systems have proven to be crucial for the in vitro maintenance of primary hepatocytes and the preservation of hepatic functional expression at a high level. A poly-(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide) matrix can recognize cells and promote liver function in a spheroid structure because of a specific galactose-asialoglycoprotein receptor interaction. Meanwhile, a fusion protein, E-cadherin-Fc, when incubated with various cells, has shown an enhancing effect on cellular viability and metabolism. Therefore, a hybrid substratum was developed for biomedical applications by using both of these materials to combine their advantages for primary hepatocyte cultures. The isolated cells showed a monolayer aggregate morphology on the coimmobilized surface and displayed higher functional expression than cells on traditional matrices. Furthermore, the hybrid system, in which the highest levels of cell adhesion and hepatocellular metabolism were achieved with the addition of 1% fetal bovine serum, showed a lower serum dependency than the collagen/gelatin-coated surface. Accordingly, this substrate may attenuate the negative effects of serum and further contribute to establishing a defined culture system for primary hepatocytes.

No MeSH data available.


Related in: MedlinePlus

The storage of glycogen (red) in primary hepatocytes on various surfaces after 1 day in culture.Notes: The surfaces include a PVLA-coated dish (A), the hybrid matrix (B), a gelatin-coated dish (C), and a collagen-coated dish (D). Scale bar: 50 μm.Abbreviation: PVLA, poly-(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide).
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f7-ijn-10-2313: The storage of glycogen (red) in primary hepatocytes on various surfaces after 1 day in culture.Notes: The surfaces include a PVLA-coated dish (A), the hybrid matrix (B), a gelatin-coated dish (C), and a collagen-coated dish (D). Scale bar: 50 μm.Abbreviation: PVLA, poly-(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide).

Mentions: In addition to their protein secreting capability, hepatocytes possess many other functionalities, such as the synthesis of biomolecules, transformation of carbohydrates, and formation of bile. Here, the functional expression of primary hepatocytes was revealed by their glycogen storage capability after 1 day (Figure 7). The hepatocytes showed an elevated level of stored glycogen (red color) on the PVLA surface (Figure 7A) and on the coimmobilized matrix (Figure 7B) with a cluster morphology, in contrast to the spreading cells on the gelatin (Figure 7C) and on the collagen surface (Figure 7D). The amount of stored glycogen was highest in cells on the PVLA matrix, although the cell number there was the smallest.


A hybrid substratum for primary hepatocyte culture that enhances hepatic functionality with low serum dependency.

Meng Q, Tao C, Qiu Z, Akaike T, Cui F, Wang X - Int J Nanomedicine (2015)

The storage of glycogen (red) in primary hepatocytes on various surfaces after 1 day in culture.Notes: The surfaces include a PVLA-coated dish (A), the hybrid matrix (B), a gelatin-coated dish (C), and a collagen-coated dish (D). Scale bar: 50 μm.Abbreviation: PVLA, poly-(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4376262&req=5

f7-ijn-10-2313: The storage of glycogen (red) in primary hepatocytes on various surfaces after 1 day in culture.Notes: The surfaces include a PVLA-coated dish (A), the hybrid matrix (B), a gelatin-coated dish (C), and a collagen-coated dish (D). Scale bar: 50 μm.Abbreviation: PVLA, poly-(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide).
Mentions: In addition to their protein secreting capability, hepatocytes possess many other functionalities, such as the synthesis of biomolecules, transformation of carbohydrates, and formation of bile. Here, the functional expression of primary hepatocytes was revealed by their glycogen storage capability after 1 day (Figure 7). The hepatocytes showed an elevated level of stored glycogen (red color) on the PVLA surface (Figure 7A) and on the coimmobilized matrix (Figure 7B) with a cluster morphology, in contrast to the spreading cells on the gelatin (Figure 7C) and on the collagen surface (Figure 7D). The amount of stored glycogen was highest in cells on the PVLA matrix, although the cell number there was the smallest.

Bottom Line: A poly-(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide) matrix can recognize cells and promote liver function in a spheroid structure because of a specific galactose-asialoglycoprotein receptor interaction.Meanwhile, a fusion protein, E-cadherin-Fc, when incubated with various cells, has shown an enhancing effect on cellular viability and metabolism.The isolated cells showed a monolayer aggregate morphology on the coimmobilized surface and displayed higher functional expression than cells on traditional matrices.

View Article: PubMed Central - PubMed

Affiliation: School of Materials Science and Engineering, Tsinghua University, Beijing, People's Republic of China ; State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, People's Republic of China ; Biomaterials Center for Regenerative Medical Engineering, Ibaraki, Japan.

ABSTRACT
Cell culture systems have proven to be crucial for the in vitro maintenance of primary hepatocytes and the preservation of hepatic functional expression at a high level. A poly-(N-p-vinylbenzyl-4-O-β-D-galactopyranosyl-D-gluconamide) matrix can recognize cells and promote liver function in a spheroid structure because of a specific galactose-asialoglycoprotein receptor interaction. Meanwhile, a fusion protein, E-cadherin-Fc, when incubated with various cells, has shown an enhancing effect on cellular viability and metabolism. Therefore, a hybrid substratum was developed for biomedical applications by using both of these materials to combine their advantages for primary hepatocyte cultures. The isolated cells showed a monolayer aggregate morphology on the coimmobilized surface and displayed higher functional expression than cells on traditional matrices. Furthermore, the hybrid system, in which the highest levels of cell adhesion and hepatocellular metabolism were achieved with the addition of 1% fetal bovine serum, showed a lower serum dependency than the collagen/gelatin-coated surface. Accordingly, this substrate may attenuate the negative effects of serum and further contribute to establishing a defined culture system for primary hepatocytes.

No MeSH data available.


Related in: MedlinePlus