Journal de la science cellulaire et de la thérapie
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Bone Marrow-Derived Mononuclear Cells Differentiate into Hepatocyte-Like Cells within Few Hrs without Fusion

Ursula Matte, Carolina Uribe Cruz, Monica Lujan López, Laura Simon, Fabiana Quoos Mayer and Roberto Giugliani

Background: Cell therapy using Bone Marrow Mononuclear Cells (BMMC) has been shown as a potential treatment for liver diseases. BMMC can act by fusion, differentiation into hepatocyte-like cells and/or secretion of paracrine factors. Here, we used encapsulated BMMC in a model of Carbon Tetrachloride (CCl4)-induced acute liver injury to study in vivo and in vitro differentiation of BMMC.

Methods: Both in vitro and in vivo studies were conducted in Wistar rats submitted to CCl4-induced acute liver injury. BMMC were isolated from Wistar rats and encapsulated in sodium alginate microcapsules. For in vivo experiments, animals received encapsulated BMMC 24 hrs after CCl4 administration and capsules were collected within 6, 24 and 48 hrs (tCCl4 group). For in vitro experiments, isolated hepatocytes from animals with CCl4-induced liver injury were co-cultured with encapsulated BMMC for 6 h (cCCl4 group). Control groups were not submitted to CCl4 administration. The content of intracellular lipid droplets in hepatocytes was used to evaluate liver injury. BMMC differentiation was assessed by RT-PCR for hepatic genes and ability to produce and secrete urea.

Results: Liver damage was confirmed in CCl4 treated animals by the presence of intracellular lipid droplets in hepatocytes and the characteristic nutmeg aspect of the liver. Retrieved encapsulated BMMC from tCCl4 group expressed hepatocyte markers, such as Cytokeratin 18 and Albumin 48 hrs after treatment. On the other hand, BMMC from cCCl4 group showed Albumin expression 6 hrs after co-culture. Urea production was increased in BMMC from cCCl4 group but not in cControl. BMMC from tControl or cControl groups did not express hepatocyte markers at any time point.

Conclusions: In this study we show that BMMC differentiate into hepatocyte-like cells in a short period of time both in vivo and in vitro. This differentiation is triggered by paracrine factors present only in injured liver.