Characterisation of Adult Liver Derived Progenitor Cells (ALDPC) and use for cell therapy and in vitro drug testing

Primary hepatocytes rapidly dedifferentiate in tissue culture and thus can be used for research purposes only for approximately 2 days.However, upon long term culture of primary hepatocytes (plate-and-wait method, [1,2]) a unique population of cells appears, we dubbed them Adult Liver Progenitor Cells (ALDPC) and are meanwhile able to generate them on a standardised basis from murine and human samples.ALDPC have a liver cell progenitor phenotype, i.e. depending on culture conditions they develop hepatic resp. cholangiocytic characteristics. Futhermore, the cells proliferate in tissue culture without obvious signs of aging/degeneration even after approx. 60 passages.

Not only to study those cells in more detail, but also to learn more about hepatocyte (de-)differentiation, -proliferation, -maturation and growth needs in general we developed a RT-qPCR panel for murine resp. human liver cells. These two panels analyse the expression of approx. 42 hepatocyte and hepatocyte progenitor specific transcripts. They are fully standardised and calibrated for state of the art RT-qPCR and easy and fast to handle–allowing comprehensive analysis of single factors in a multifactorial system.The use of the panels enbled us to

i) investigate primary, fresh hepatocyte dedifferentiation over time,

ii) refine and standardise culture conditions for hepatocytes and ALDPC,

iii) define requirements for 1) optimal proliferation and 2) in vitro rematuration towards a highly metabolical active, non proliferative phenotype of ALDPC,

iv) define conditions for priming of ALDPCs for cell transplantation and functional engraftment in appropriate mouse models.

Data will be presented describing the usefulness of above described RT-qPCR panels and demonstrating the potentials of ALDPCs not only for use in liver cell therapy but also for in vitro drug resp. drug toxicity testings.

Literature:

[1] Strick-Marchand H, Weiss MC.Inducible differentiation and morphogenesis of bipotential liver cell lines from wild-type mouse embryos.Hepatology. 2002;36:794-804. [2] Fougère-Deschatrette C, Imaizumi-Scherrer T, Strick-Marchand H, Morosan S, Charneau P, Kremsdorf D, Faust DM, Weiss MC.Plasticity of hepatic cell differentiation: bipotential adult mouse liver clonal cell lines competent to differentiate in vitro and in vivo.Stem Cells. 2006;24:2098-109.

cell therapy - drug screening - hepatocyte progenitors - maturation - proliferation - qPCR