Decellularized　liver　matrix　(DLM)　hold　great　potential　for　reconstructing　functional　hepatic-like　tissue　(HLT)　based　on　reseeding　of　hepatocytes　or　stem　cells,　but　the　shortage　of　liver　donors　is　still　an　obstacle　for　potential　application.　Therefore,　an　appropriate　alternative　scaffold　is　needed　to　expand　the　donor　pool.　In　this　study,　we　explored　the　effectiveness　of　decellularized　spleen　matrix　(DSM)　for　culturing　of　bone　marrow　mesenchymal　stem　cells　(BMSCs),　and　promoting　differentiation　into　hepatic-like　cells.Rats＇　spleen　were　harvested　for　DSM　preparation　by　freezing/thawing　and　perfusion　procedure.　Then　the　mesenchymal　stem　cells　derived　from　rat　bone　marrow　were　reseeded　into　DSM　for　dynamic　culture　and　hepatic　differentiation　by　a　defined　induction　protocol.The　research　found　that　DSM　preserved　a　3-dimensional　porous　architecture,　with　native　extracellular　matrix　and　vascular　network　which　was　similar　to　DLM.　The　reseeded　BMSCs　in　DSM　differentiated　into　functional　hepatocyte-like　cells,　evidenced　by　cytomorphology　change,　expression　of　hepatic-associated　genes　and　protein　markers,　glycogen　storage,　and　indocyanine　green　uptake.　The　albumin　production　(2.74±0.42　vs.　2.07±0.28　pg/cell/day)　and　urea　concentration　(75.92±15.64　vs.　52.07±11.46　pg/cell/day)　in　DSM　group　were　remarkably　higher　than　tissue　culture　flasks　(TCF)　group　over　the　same　differentiation　period,　P＜　0.05.This　present　study　demonstrated　that　DSM　might　have　considerable　potential　in　fabricating　hepatic-like　tissue,　particularly　because　it　can　facilitate　hepatic　differentiation　of　BMSCs　which　exhibited　higher　level　and　more　stable　functions.