title: Morphological and functional analysis of rat hepatocyte spheroids generated on poly(L-lactic acid) polymer in a pulsatile flow bioreactor.
abstract: Liver neo-tissue suitable for transplantation has not been established. Primary rat hepatocytes were cultured on three-dimensional biodegradable polymer matrices in a pulsatile flow bioreactor with the intention of inducing tissue formation and improving cell survival. Functional and structural analysis of the hepatocytes forming liver neo-tissue was performed. Biodegradable poly(L-lactic acid) (PLLA) polymer discs were seeded with 4 x 10(6) primary rat hepatocytes each, were exposed to a pulsatile medium flow of 24 mL/min for 1, 2, 4, or 6 days and were investigated for monoethylglycinexylidine (MEGX) formation, ammonia detoxification, Cytokeratin 18 (CK18) expression, and preserved glycogen storage. Fine structural details were obtained using scanning and transmission electron microscopy. Spheroids of viable hepatocytes were formed. MEGX-specific production was maintained and ammonia removal capacity remained high during the entire flow-culture period of 6 days. CK18 distribution was normal. Periodic-acid- Schiff reaction demonstrated homogenous glycogen storage. The hepatocytes reassembled to form intercellular junctions and bile canaliculi. Functional and morphological analysis of rat hepatocytes forming spheroids in a pulsatile flow bioreactor indicated preserved and intact hepatocyte morphology and specific function. Pulsatile flow culture on PLLA scaffolds is a promising new method of hepatic tissue engineering leading to liver neo-tissue formation.
authors.affiliation: Department of Hepatobiliary Surgery and Visceral Transplantation, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Török
authors.forename: Christian
authors.initials: C
authors.lastname: Vogel
authors.forename: Marc
authors.initials: M
authors.lastname: Lütgehetmann
authors.forename: Peter X
authors.initials: PX
authors.lastname: Ma
authors.forename: Maura
authors.initials: M
authors.lastname: Dandri
authors.forename: Joerg
authors.initials: J
authors.lastname: Petersen
authors.forename: Martin R
authors.initials: MR
authors.lastname: Burda
authors.forename: Klaus
authors.initials: K
authors.lastname: Siebert
authors.forename: Jochen
authors.initials: J
authors.lastname: Düllmann
authors.forename: Xavier
authors.initials: X
authors.lastname: Rogiers
authors.forename: Joerg-M
authors.initials: JM
authors.lastname: Pollok
mesh_terms: D000818:Animals; D001672:Biocompatible Materials; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002478:Cells, Cultured; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology / Q000637:transplantation; D019344:Lactic Acid; D008099:Liver / Q000166:cytology* / Q000502:physiology; D019164:Liver, Artificial; D008297:Male; D011091:Polyesters; D011108:Polymers; D011673:Pulsatile Flow; D051381:Rats; D011917:Rats, Inbred Lew; D018874:Spheroids, Cellular / Q000166:cytology* / Q000502:physiology / Q000637:transplantation; D023822:Tissue Engineering
chemical_list: D001672:Biocompatible Materials; D011091:Polyesters; D011108:Polymers; D019344:Lactic Acid; C033616:poly(lactide)