Rat hepatocyte aggregate formation on discrete aligned nanofibers of type-I collagen-coated poly(L-lactic acid).
Primary hepatocytes cultured in three dimensional tissue constructs composed of multicellular aggregates maintain normal differentiated cellular function in vitro while cultured monolayers do not. Here, we report a technique to induce hepatocyte aggregate formation using type-I collagen-coated poly(L-lactic acid) (PLLA) discrete aligned nanofibers (disAFs) by providing limited cell-substrate adhesion strength and restricting cell migration to uniaxial movement. Kinetics of aggregate formation, morphology and biochemical activities of rat hepatocyte aggregates were tested over a 15 day culture period. Evidence was provided that physical cues from disAFs quickly induced the formation of aggregates. After 3 days in culture, 88.3% of free hepatocytes on disAFs were incorporated into aggregates with an average diameter of 61 +/- 18 microm. Hepatocyte aggregates formed on disAFs displayed excellent cell retention, cell activity and stable functional expression in terms of albumin secretion, urea synthesis and phase I and II (CYP1A and UGT) metabolic enzyme activity compared to monolayer culture of hepatocytes on tissue culture plastic (TCP) with type-I collagen as well as on meshes of type-I collagen-coated PLLA random nanofibers (meshRFs). These results suggest that disAFs may be a suitable method to maintain large-scale hepatic cultures with high activity for tissue engineering research and potential therapeutic applications, such as bioartificial liver devices.
Feng
Zhang-Qi
ZQ
State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China.
Chu
Xue-Hui
XH
Huang
Ning-Ping
NP
Leach
Michelle K
MK
Wang
Gan
G
Wang
Yi-Chun
YC
Ding
Yi-Tao
YT
Gu
Zhong-Ze
ZZ
D000418:Albumins / Q000378:metabolism; D000818:Animals; D024042:Collagen Type I; D005456:Fluorescent Dyes; D022781:Hepatocytes / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D019344:Lactic Acid; D008297:Male; D008855:Microscopy, Electron, Scanning; D057139:Nanofibers; D011091:Polyesters; D011108:Polymers; D051381:Rats; D017207:Rats, Sprague-Dawley; D014508:Urea / Q000378:metabolism
D000418:Albumins; D024042:Collagen Type I; D005456:Fluorescent Dyes; D011091:Polyesters; D011108:Polymers; D019344:Lactic Acid; C033616:poly(lactide); D014508:Urea