Synthetic sandwich culture of 3D hepatocyte monolayer.
The sandwich culture of hepatocytes, between double layers of extra-cellular matrix (ECM), is a well-established in vitro model for re-establishing hepatic polarity and maintaining differentiated functions. Applications of the ECM-based sandwich culture are limited by the mass transfer barriers induced by the top gelled ECM layer, complex molecular composition of ECM with batch-to-batch variation and uncontrollable coating of the ECM double layers. We have addressed these limitations of the ECM-based sandwich culture by developing an 'ECM-free' synthetic sandwich culture, which is constructed by sandwiching a 3D hepatocyte monolayer between a glycine-arginine-glycine-aspatic acid-serine (GRGDS)-modified polyethylene terephthalate (PET) track-etched membrane (top support) and a galactosylated PET film (bottom substratum). The bioactive top support and bottom substratum in the synthetic sandwich culture substituted for the functionalities of the ECM in the ECM-based sandwich culture with further improvement in mass transfer and optimal material properties. The 3D hepatocyte monolayer in the synthetic sandwich culture exhibited a similar process of hepatic polarity formation, better cell-cell interaction and improved differentiated functions over 14-day culture compared to the hepatocytes in collagen sandwich culture. The novel 3D hepatocyte monolayer sandwich culture using bioactive synthetic materials may readily replace the ECM-based sandwich culture for liver tissue engineering applications, such as drug metabolism/toxicity testing and hepatocyte-based bioreactors.
Du
Yanan
Y
Institute of Bioengineering and Nanotechnology, A STAR, Singapore 138669, Singapore.
Han
Rongbin
R
Wen
Feng
F
Ng San San
Susanne
S
Xia
Lei
L
Wohland
Thorsten
T
Leo
Hwa Liang
HL
Yu
Hanry
H
D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D016764:Cell Polarity; D048430:Cell Shape; D002478:Cells, Cultured; D003094:Collagen; D022781:Hepatocytes / Q000166:cytology*; D008297:Male; D008855:Microscopy, Electron, Scanning; D011093:Polyethylene Terephthalates; D051381:Rats; D017208:Rats, Wistar
D011093:Polyethylene Terephthalates; D003094:Collagen