Enhanced hepatic differentiation of rat bone marrow-derived mesenchymal stem cells in spheroidal aggregate culture on a decellularized liver scaffold.
Enhanced hepatic differentiation of rat bone marrow-derived mesenchymal stem cells in spheroidal aggregate culture on a decellularized liver scaffold.
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DOI：
10.3892/ijmm.2016.2638
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发表时间：
2016-08
影响因子：
5.4
通讯作者：
Bu H
中科院分区：
文献类型：
--
作者：
Bao J;Wu Q;Wang Y;Li Y;Li L;Chen F;Wu X;Xie M;Bu H
在本研究中，我们的目的是确定聚集培养物和脱细胞肝脏支架（DLS）的结合是否可以促进小鼠骨髓来源的间充质干细胞（BM-MSC）在体外向高产的成熟肝细胞分化。研究了四种分化培养方法[单细胞(2D)、球状体(3D)、2D + DLS 和3D + DLS]。为了确定 MSC 的分化阶段，进行了肝细胞基因的 RT-qPCR、肝细胞标记物的免疫染色和功能分析。与其他组相比，细胞外基质（ECM）上由BM-MSC球体分化而来的肝细胞样细胞表现出更强烈的储存糖原染色、更高水平的尿素生物合成和白蛋白分泌以及更高的肝细胞特异性基因表达。我们的结果表明，DLS 与球状聚集体培养相结合可作为促进 BM-MSC 肝脏成熟的有效方法，并可能在基于干细胞的肝脏再生医学中具有未来的应用。
In the present study, we aimed to determine whether the combination of aggregate culture and decellularized liver scaffolds (DLSs) promoted the hepatic differentiation of murine bone marrow-derived mesenchymal stem cells (BM-MSCs) into high yields of mature hepatocytes in vitro. Four culturing methods for differentiation [single cell (2D), spheroids (3D), 2D + DLS and 3D + DLS] were studied. To determine the differentiation stages of the MSCs, RT-qPCR of the hepatocyte genes, immunostaining of hepatocyte markers, and functional analyses were all performed. Compared with the other groups, hepatocyte-like cells which differentiated from BM-MSC spheroids on extracellular matrix (ECM) exhibited more intensive staining of stored glycogen, an elevated level of urea biosynthesis and albumin secretion as well as the higher expression of hepatocyte-specific genes. Our results indicated that DLSs combined with spheroidal aggregate culture may be used as an effective method to facilitate the hepatic maturation of BM-MSCs and may have future applications in stem cell-based liver regenerative medicine.