Abstract
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.
Highlights
Bone Marrow Mononuclear Cells (BMMC) have emerged as potential candidates for cell therapy due to their ease of use
Recent clinical trials have demonstrated that transplantation of these cells or their fractions improves the condition of patients with cirrhosis [3,4,5], as well as improve liver function in animal models of cirrhosis [6]
In vitro experiments Since encapsulated BMMC showed the expression of hepatocyte markers 48 hrs after implantation in animals with CCl4-induced acute liver injury, we tried to replicate the same model in vitro in order to determine when this reprogramming takes place
Summary
Bone Marrow Mononuclear Cells (BMMC) have emerged as potential candidates for cell therapy due to their ease of use. Recent clinical trials have demonstrated that transplantation of these cells or their fractions improves the condition of patients with cirrhosis [3,4,5], as well as improve liver function in animal models of cirrhosis [6]. Many groups have developed protocols for the in vitro differentiation of bone marrow-derived cells into hepatocyte-like cells [19,20,21,22,23,24]. Bone marrow mononuclear fraction is used for in vivo transplantation, in vitro protocols usually work with Mesenchymal Stem Cells (MSC) [19,23,24,25]. Cell therapy using Bone Marrow Mononuclear Cells (BMMC) has been shown as a potential treatment for liver diseases. 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
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
