[Integrative Biology] Harnessing Endogenous Signals from Hepatocytes Using a Low Volume Multi-Well P
Hepatocytes are highly differentiated epithelial cells that lose their phenotype and function when removed from the in vivo environment. Given the importance of hepatic cultures for drug toxicity, bioartificial liver assist devices and basic biology studies, considerable efforts have been focused on maintenance of hepatic function in vitro. The methods used to date include co-cultivation of hepatocytes with stromal cells, organizing these cells into spheroids and imbedding them into bioactive gels. Our team has recently demonstrated that primary rat hepatocytes confined to microfluidic channels in the absence of convection maintained epithelial phenotype through upregulation of endogenous signals including hepatocyte growth factor (HGF). The objective of the present study was to transition from microfluidic devices which are somewhat specialized and challenging to use and towards low volume multiwell plates ubiquitous in the biology laboratories. Using a combination of 3D printing and micromolding we have constructed inserts that could be placed into standard 12-well plates and could be used to create low volume culture conditions under which primary hepatocytes maintained differentiated phenotype. This phenotype enhancement was confirmed by assays including albumin synthesis and expression. Importantly we confirmed upregulation of HGF inside the low volume culture plates and demonstrated that inhibition of HGF signaling degraded hepatic phenotype in our cell culture platform. Overall, this study outlines a new cell culture system that leverages low volume effects of microfluidic channels in a multi-well plate format. Beyond hepatocytes, such system may be of use in maintenance of other difficult-to-culture cells including stem cells and primary cancer cells.
Pantea Gheibi, Kyung Jin Son, Gulnaz Stybayeva and Alexander Revzin
Integr. Biol., 2017, Accepted Manuscript