Enhanced hepatic differentiation of rat bone marrow-derived mesenchymal stem cells in spheroidal aggregate culture on a decellularized liver scaffold.
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.
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Ji
J
Bao
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Qiong
Q
Wu
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Yujia
Y
Wang
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Yi
Y
Li
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Li
L
Li
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Fei
F
Chen
Department of General Surgery, Yibin City First People's Hospital, Yibin, Sichuan 644000, P.R. China.
Xiujuan
X
Wu
Department of General Surgery, Yibin City First People's Hospital, Yibin, Sichuan 644000, P.R. China.
Mingjun
M
Xie
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Hong
H
Bu
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002449:Cell Aggregation; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D018874:Spheroids, Cellular / Q000166:cytology*; D013552:Swine; D013556:Swine, Miniature; D054457:Tissue Scaffolds / Q000737:chemistry*
D015415:Biomarkers