title: Long-term stability of primary rat hepatocytes in micropatterned cocultures.
abstract: Primary hepatocytes display functional and structural instability in standard monoculture systems. We have previously developed a model in which primary hepatocytes are organized in domains of empirically optimized dimensions and surrounded by murine embryonic fibroblasts (HepatoPac™). Here, we assess the long-term phenotype of freshly isolated and cryopreserved rat hepatocytes in a 96-well HepatoPac format. The viability, cell polarity (actin microfilaments, bile canaliculi), and functions (albumin, urea, Phase I/II enzymes, transporters) of fresh and cryopreserved rat hepatocytes were retained in HepatoPac at similar levels for at least 4 weeks as opposed to rapidly declining over 5 days in collagen/Matrigel™ sandwich cultures. Pulse or continuous exposure of rat HepatoPac to GW-7647, a selective agonist of PPARα, caused reproducible induction of CYP4A1 and 3-hydroxy-3-methylglutaryl-CoA synthase over 4 weeks. In conclusion, rat HepatoPac in a 96-well format can be used for chronic dosing of highly functional hepatocytes and assessment of perturbed hepatocellular pathways.
authors: Hepregen Corporation, Suite 1500, Medford, MA 2155, USA.
authors: Okechukwu
authors: O
authors: Ukairo
authors: Chitra
authors: C
authors: Kanchagar
authors: Amanda
authors: A
authors: Moore
authors: Julianne
authors: J
authors: Shi
authors: Jeannemarie
authors: J
authors: Gaffney
authors: Simon
authors: S
authors: Aoyama
authors: Kelly
authors: K
authors: Rose
authors: Stacy
authors: S
authors: Krzyzewski
authors: Jack
authors: J
authors: McGeehan
authors: Melvin E
authors: ME
authors: Andersen
authors: Salman R
authors: SR
authors: Khetani
authors: Edward L
authors: EL
authors: Lecluyse
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D016764:Cell Polarity / Q000235:genetics; D018920:Coculture Techniques / Q000379:methods*; D015925:Cryopreservation; D005347:Fibroblasts / Q000166:cytology*; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D051379:Mice; D051381:Rats