Reconstruction of hepatic stellate cell-incorporated liver capillary structures in small hepatocyte tri-culture using microporous membranes

Junichi Kasuya, Ryo Sudo, Genta Masuda, Toshihiro Mitaka, Mariko Ikeda, Kazuo Tanishita

研究成果: Article

6 引用 (Scopus)

抄録

In liver sinusoids, hepatic stellate cells (HSCs) locate the outer surface of microvessels to form a functional unit with endothelia and hepatocytes. To reconstruct functional liver tissue in vitro, formation of the HSC-incorporated sinusoidal structure is essential. We previously demonstrated capillary formation of endothelial cells (ECs) in tri-culture, where a polyethylene terephthalate (PET) microporous membrane was intercalated between the ECs and hepatic organoids composed of small hepatocytes (SHs), i.e. hepatic progenitor cells, and HSCs. However, the high thickness and low porosity of the membranes limited heterotypic cell-cell interactions, which are essential to form HSC-EC hybrid structures. Here, we focused on the effective use of the thin and highly porous poly( d, l-lactide-co-glycolide) (PLGA) microporous membranes in SH-HSC-EC tri-culture to reconstruct the HSC-incorporated liver capillary structures in vitro. First, the formation of EC capillary-like structures was induced on Matrigel-coated PLGA microporous membranes. Next, the membranes were stacked on hepatic organoids composed of small SHs and HSCs. When the pore size and porosity of the membranes were optimized, HSCs selectively migrated to the EC capillary-like structures. This process was mediated in part by platelet-derived growth factor (PDGF) signalling. In addition, the HSCs were located along the outer surface of the EC capillary-like structures with their long cytoplasmic processes. In the HSC-incorporated capillary tissues, SHs acquired high levels of differentiated functions, compared to those without ECs. This model will provide a basis for the construction of functional, thick, vascularized liver tissues in vitro.

元の言語English
ページ(範囲)247-256
ページ数10
ジャーナルJournal of Tissue Engineering and Regenerative Medicine
9
発行部数3
DOI
出版物ステータスPublished - 2015 3 1
外部発表Yes

Fingerprint

Hepatic Stellate Cells
Liver
Endothelial cells
Hepatocytes
Membranes
Endothelial Cells
Organoids
Porosity
Tissue
Cell culture
Polyethylene Terephthalates
Platelet-Derived Growth Factor
Platelets
Microvessels
Cell Communication
Polyethylene terephthalates
Pore size
Endothelium
Stem Cells
Cell Culture Techniques

ASJC Scopus subject areas

  • Biomedical Engineering
  • Medicine (miscellaneous)
  • Biomaterials

これを引用

Reconstruction of hepatic stellate cell-incorporated liver capillary structures in small hepatocyte tri-culture using microporous membranes. / Kasuya, Junichi; Sudo, Ryo; Masuda, Genta; Mitaka, Toshihiro; Ikeda, Mariko; Tanishita, Kazuo.

:: Journal of Tissue Engineering and Regenerative Medicine, 巻 9, 番号 3, 01.03.2015, p. 247-256.

研究成果: Article

Kasuya, Junichi ; Sudo, Ryo ; Masuda, Genta ; Mitaka, Toshihiro ; Ikeda, Mariko ; Tanishita, Kazuo. / Reconstruction of hepatic stellate cell-incorporated liver capillary structures in small hepatocyte tri-culture using microporous membranes. :: Journal of Tissue Engineering and Regenerative Medicine. 2015 ; 巻 9, 番号 3. pp. 247-256.
@article{e965c9bbd60b47a1a267af5812b279b9,
title = "Reconstruction of hepatic stellate cell-incorporated liver capillary structures in small hepatocyte tri-culture using microporous membranes",
abstract = "In liver sinusoids, hepatic stellate cells (HSCs) locate the outer surface of microvessels to form a functional unit with endothelia and hepatocytes. To reconstruct functional liver tissue in vitro, formation of the HSC-incorporated sinusoidal structure is essential. We previously demonstrated capillary formation of endothelial cells (ECs) in tri-culture, where a polyethylene terephthalate (PET) microporous membrane was intercalated between the ECs and hepatic organoids composed of small hepatocytes (SHs), i.e. hepatic progenitor cells, and HSCs. However, the high thickness and low porosity of the membranes limited heterotypic cell-cell interactions, which are essential to form HSC-EC hybrid structures. Here, we focused on the effective use of the thin and highly porous poly( d, l-lactide-co-glycolide) (PLGA) microporous membranes in SH-HSC-EC tri-culture to reconstruct the HSC-incorporated liver capillary structures in vitro. First, the formation of EC capillary-like structures was induced on Matrigel-coated PLGA microporous membranes. Next, the membranes were stacked on hepatic organoids composed of small SHs and HSCs. When the pore size and porosity of the membranes were optimized, HSCs selectively migrated to the EC capillary-like structures. This process was mediated in part by platelet-derived growth factor (PDGF) signalling. In addition, the HSCs were located along the outer surface of the EC capillary-like structures with their long cytoplasmic processes. In the HSC-incorporated capillary tissues, SHs acquired high levels of differentiated functions, compared to those without ECs. This model will provide a basis for the construction of functional, thick, vascularized liver tissues in vitro.",
keywords = "Endothelial cell, Hepatic stellate cell, Liver sinusoid, Microporous membrane, Poly(d, l-lactide-co-glycolide), Small hepatocyte, Three-dimensional, Tri-culture",
author = "Junichi Kasuya and Ryo Sudo and Genta Masuda and Toshihiro Mitaka and Mariko Ikeda and Kazuo Tanishita",
year = "2015",
month = "3",
day = "1",
doi = "10.1002/term.1630",
language = "English",
volume = "9",
pages = "247--256",
journal = "Journal of Tissue Engineering and Regenerative Medicine",
issn = "1932-6254",
publisher = "John Wiley and Sons Ltd",
number = "3",

}

TY - JOUR

T1 - Reconstruction of hepatic stellate cell-incorporated liver capillary structures in small hepatocyte tri-culture using microporous membranes

AU - Kasuya, Junichi

AU - Sudo, Ryo

AU - Masuda, Genta

AU - Mitaka, Toshihiro

AU - Ikeda, Mariko

AU - Tanishita, Kazuo

PY - 2015/3/1

Y1 - 2015/3/1

N2 - In liver sinusoids, hepatic stellate cells (HSCs) locate the outer surface of microvessels to form a functional unit with endothelia and hepatocytes. To reconstruct functional liver tissue in vitro, formation of the HSC-incorporated sinusoidal structure is essential. We previously demonstrated capillary formation of endothelial cells (ECs) in tri-culture, where a polyethylene terephthalate (PET) microporous membrane was intercalated between the ECs and hepatic organoids composed of small hepatocytes (SHs), i.e. hepatic progenitor cells, and HSCs. However, the high thickness and low porosity of the membranes limited heterotypic cell-cell interactions, which are essential to form HSC-EC hybrid structures. Here, we focused on the effective use of the thin and highly porous poly( d, l-lactide-co-glycolide) (PLGA) microporous membranes in SH-HSC-EC tri-culture to reconstruct the HSC-incorporated liver capillary structures in vitro. First, the formation of EC capillary-like structures was induced on Matrigel-coated PLGA microporous membranes. Next, the membranes were stacked on hepatic organoids composed of small SHs and HSCs. When the pore size and porosity of the membranes were optimized, HSCs selectively migrated to the EC capillary-like structures. This process was mediated in part by platelet-derived growth factor (PDGF) signalling. In addition, the HSCs were located along the outer surface of the EC capillary-like structures with their long cytoplasmic processes. In the HSC-incorporated capillary tissues, SHs acquired high levels of differentiated functions, compared to those without ECs. This model will provide a basis for the construction of functional, thick, vascularized liver tissues in vitro.

AB - In liver sinusoids, hepatic stellate cells (HSCs) locate the outer surface of microvessels to form a functional unit with endothelia and hepatocytes. To reconstruct functional liver tissue in vitro, formation of the HSC-incorporated sinusoidal structure is essential. We previously demonstrated capillary formation of endothelial cells (ECs) in tri-culture, where a polyethylene terephthalate (PET) microporous membrane was intercalated between the ECs and hepatic organoids composed of small hepatocytes (SHs), i.e. hepatic progenitor cells, and HSCs. However, the high thickness and low porosity of the membranes limited heterotypic cell-cell interactions, which are essential to form HSC-EC hybrid structures. Here, we focused on the effective use of the thin and highly porous poly( d, l-lactide-co-glycolide) (PLGA) microporous membranes in SH-HSC-EC tri-culture to reconstruct the HSC-incorporated liver capillary structures in vitro. First, the formation of EC capillary-like structures was induced on Matrigel-coated PLGA microporous membranes. Next, the membranes were stacked on hepatic organoids composed of small SHs and HSCs. When the pore size and porosity of the membranes were optimized, HSCs selectively migrated to the EC capillary-like structures. This process was mediated in part by platelet-derived growth factor (PDGF) signalling. In addition, the HSCs were located along the outer surface of the EC capillary-like structures with their long cytoplasmic processes. In the HSC-incorporated capillary tissues, SHs acquired high levels of differentiated functions, compared to those without ECs. This model will provide a basis for the construction of functional, thick, vascularized liver tissues in vitro.

KW - Endothelial cell

KW - Hepatic stellate cell

KW - Liver sinusoid

KW - Microporous membrane

KW - Poly(d, l-lactide-co-glycolide)

KW - Small hepatocyte

KW - Three-dimensional

KW - Tri-culture

UR - http://www.scopus.com/inward/record.url?scp=84925677456&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84925677456&partnerID=8YFLogxK

U2 - 10.1002/term.1630

DO - 10.1002/term.1630

M3 - Article

C2 - 23086892

AN - SCOPUS:84925677456

VL - 9

SP - 247

EP - 256

JO - Journal of Tissue Engineering and Regenerative Medicine

JF - Journal of Tissue Engineering and Regenerative Medicine

SN - 1932-6254

IS - 3

ER -