Effect of a hepatocyte growth factor/heparin-immobilized collagen system on albumin synthesis and spheroid formation by hepatocytes.
A hepatocyte growth factor (HGF)/heparin-immobilized collagen system was used as a synthetic extracellular matrix for hepatocyte culture. The albumin synthesis, nucleus numbers and morphology of the hepatocytes were determined separately to evaluate the hepatocyte number and hepatocyte-specific function under this system. The benefits of the HGF/heparin-immobilized collagen system for hepatocyte culture were confirmed by three types of culture methods in vitro, namely 2D film cultures, 2D gel cultures and 3D gel cultures. In 2D collagen film cultures, hepatocytes exhibited the highest albumin synthesis (1.42 microg/well/day) in HGF/heparin-immobilized collagen films at 7 days of culture. Heparin inhibited hepatocyte adhesion while HGF promoted hepatocyte migration, and spheroid formation was easily detected in HGF/heparin-immobilized collagen films. In 2D collagen gel cultures, albumin synthesis of around 15 microg/well/day was detected and maintained for more than 18 days on HGF/heparin-immobilized collagen gels. Similar findings were obtained in 3D HGF/heparin-immobilized collagen gel cultures, which exhibited albumin synthesis of up to 30 microg/well/day. The albumin synthesis by hepatocytes was two-fold higher in 3D gel cultures compared with 2D gel cultures, and was maintained for over 2 weeks compared with 2D film cultures using the HGF/heparin-immobilized collagen system. Taken together, the HGF/heparin-immobilized collagen system was effective for albumin synthesis by hepatocytes in both 2D film cultures and 3D gel cultures, and therefore shows good potential for tissue engineering use.
Hou
Yung-Te
YT
Department of Chemical Engineering, Faculty of Engineering, Graduate School, Kyushu University, 744 Motooka, Nishi-Ku, Fukuoka, 819-0395, Japan.
Ijima
Hiroyuki
H
Matsumoto
Shunichi
S
Kubo
Takafumi
T
Takei
Takayuki
T
Sakai
Shinji
S
Kawakami
Koei
K
D000818:Animals; D000925:Anticoagulants / Q000008:administration & dosage / Q000737:chemistry; D049108:Cell Enlargement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000008:administration & dosage* / Q000737:chemistry; D006493:Heparin / Q000008:administration & dosage* / Q000737:chemistry; D017228:Hepatocyte Growth Factor / Q000008:administration & dosage* / Q000737:chemistry; D022781:Hepatocytes / Q000187:drug effects / Q000502:physiology*; D019164:Liver, Artificial; D008297:Male; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D012709:Serum Albumin / Q000096:biosynthesis*; D018874:Spheroids, Cellular / Q000187:drug effects / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods
D000925:Anticoagulants; D012709:Serum Albumin; D017228:Hepatocyte Growth Factor; D006493:Heparin; D003094:Collagen