Microfabricated silicon nitride membranes for hepatocyte sandwich culture.
We have developed a hepatocyte sandwich culture with improved mass transport properties based on ultra-thin microfabricated porous silicon nitride (Si(3)N(4)) membranes. The dimensions and uniformity of the membrane pores can be configurable, which confers more control over the mass transport. Instead of collagen gels used in conventional sandwich culture, we utilized galactose ligands immobilized on the Si(3)N(4) membranes to support hepatocyte attachment and function in the sandwich culture. Diffusion studies using FITC-dextrans confirmed that mass transport of the microfabricated Si(3)N(4) membrane based sandwich was significantly better than conventional collagen gel sandwich and can be configured by varying the porosity of the Si(3)N(4) membrane. Hepatocytes cultured in the microfabricated Si(3)N(4) membrane based sandwich culture exhibited earlier apical repolarization and biliary excretion, improved differentiated functions and enhanced drug sensitivity compared to hepatocytes cultured in a collagen gel sandwich. The Si(3)N(4) membrane based sandwich culture allows for a systematic optimization of the mass transport properties of hepatocyte culture by changing the pore size and inter-pore distance. This will enable more effective drug testing applications where optimal mass transport is required for hepatocyte function maintenance and drug accessibility.
Zhang
Shufang
S
Institute of Bioengineering and Nanotechnology, A*STAR, The Nanos #04-01, 31 Biopolis Way, Singapore 138669, Singapore.
Xia
Lei
L
Kang
Chiang Huen
CH
Xiao
Guangfa
G
Ong
Siew Min
SM
Toh
Yi Chin
YC
Leo
Hwa Liang
HL
van Noort
Danny
D
Kan
Shyi Herng
SH
Tang
Hui Huan
HH
Yu
Hanry
H
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D001692:Biological Transport; D018929:Cell Culture Techniques / Q000379:methods*; D016764:Cell Polarity; D048430:Cell Shape; D002478:Cells, Cultured; D005690:Galactose / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D008297:Male; D008422:Materials Testing; D008567:Membranes, Artificial; D016062:Porosity; D051381:Rats; D017208:Rats, Wistar; D017655:Silicon Compounds / Q000737:chemistry*
D001672:Biocompatible Materials; D008567:Membranes, Artificial; D017655:Silicon Compounds; C032734:silicon nitride; D005690:Galactose