2005 Abstracts: Functional Comparison of Bone Marrow Derived Liver Stem Cells: Selection Strategy for Cell Based Therapy
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Functional Comparison of Bone Marrow Derived Liver Stem Cells: Selection Strategy for Cell Based Therapy
Daniel Inderbitzin, University Hospital Bern, Bern, Bern, Switzerland; Itzhak Avital, Department of Surgery, New York, NY; Beat Gloor, Adrian Keogh, Daniel Candinas, University Hospital Bern, Bern, Bern, Switzerland
Background: Liver progenitor cells can be isolated from adult bone marrow by different methods. Two subpopulations of bone marrow cells were obtained either from the non-adherent cell fraction after a panning procedure on a polystyrene surface or by a two-step immuno-isolation procedure (Beta-2-Microglobulin negative/Thy-1 positive selection). To determine the growth characteristics and the individual liver specific metabolic capacity, the bone marrow cells were cultured under different conditions.
Methods: The two subpopulations were cultured either on a polystyrene dish or on a layer of matrigel, in a 1:1 mixture of Dulbecco's modified Eagle medium and Ham's medium F12 supplemented with hepatocyte growth factor or in small hepatocyte media and seeded in high (100'000 cells) or low (10'000 cells) density for 12 days. Real-time PCR on the Taqman system (Applied Biosystems®) was performed according to standard protocols for 18S rRNA, albumin, Mrp1, and Mrp2. Urea formation and albumin secretion was determined by a colorimetric assay and a sandwich ELISA in the culture media and standardized by 18S rRNA content. Results: Both cell isolation procedures yielded albumin and multidrug resistance associated protein (Mrp1) positive cells. The Beta-2-Microglobulin negative/Thy-1 positive subpopulation of bone marrow cells cultured on matrigel in small hepatocyte media produced significantly more urea from ammonia and also secreted a superior amount of albumin in the culture media when compared to the panned cell fraction (p<0.001). Conclusions: The role of the Beta-2-Microglobulin negative/Thy-1 positive subpopulation of the bone marrow requires further investigation and could prove to be valuable for the development of novel cell based treatment strategies for congenital or acquired liver diseases.
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