Rat embryonic liver cell expansion and differentiation on NH3 plasma-grafted PEEK-WC-PU membranes.
Biomaterials can potentially influence stem and progenitor cell proliferation and differentiation in both a positive and a negative way. Herein, we report on the expansion and differentiation of rat embryonic (E17) liver (RLC-18) cells on new bioactive membrane made of PEEK-WC-PU, whose surface was grafted with nitrogen functionalities by means of NH(3) glow discharges. The performance of the developed membrane was evaluated by analyzing the expression of the liver specific functions of cells cultured in a 6-well gas-permeable bioreactor. It was found that native and NH(3) plasma-grafted PEEK-WC-PU membranes enabled expansion of liver cells in the bioreactor. Liver embryonic cells on the membranes exhibited higher functional activities compared to those cultured on conventional culture dishes as demonstrated by higher albumin and urea production. They showed gene expression of alpha-fetoprotein and albumin in a time-dependent manner of the hepatic differentiation process. LDH assay and SEM analyses revealed that a high number of viable liver stem cells attached to the membranes. Unexpectedly, liver progenitors cultured on membranes had higher telomerase activity than ones in the plates, preventing cell senescence. Thus, membranes are able to sustain in vitro the same in vivo liver functions and to allow the expansion of progenitor cells.
Pavlica
Sanja
S
Department of Cell Technologies and Applied Stem Cell Biology, Biomedical-Biotechnological Center, Medical Faculty, University Leipzig, Deutscher Platz 5, Leipzig 04103, Germany. sanja.pavlica@bbz.uni-leipzig.de
Piscioneri
Antonella
A
Peinemann
Frank
F
Keller
Mario
M
Milosevic
Javorina
J
Staeudte
Andrea
A
Heilmann
Andreas
A
Schulz-Siegmund
Michaela
M
Laera
Stefania
S
Favia
Pietro
P
De Bartolo
Loredana
L
Bader
Augustinus
A
D000641:Ammonia / Q000494:pharmacology*; D000818:Animals; D001577:Benzophenones; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D007659:Ketones / Q000494:pharmacology*; D008099:Liver / Q000166:cytology* / Q000196:embryology* / Q000378:metabolism / Q000648:ultrastructure; D008567:Membranes, Artificial; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011108:Polymers; D011140:Polyurethanes / Q000494:pharmacology*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000378:metabolism
D001577:Benzophenones; D007659:Ketones; D008567:Membranes, Artificial; D011108:Polymers; D011140:Polyurethanes; C063834:polyetheretherketone; D011092:Polyethylene Glycols; D000641:Ammonia; D019098:Telomerase