Continuous zebularine treatment enhances hepatic differentiation of mesenchymal stem cells under liver-specific factors induction in vitro

Life Sciences ◽  
2018 ◽  
Vol 215 ◽  
pp. 57-63
Author(s):  
Juan Chen ◽  
Qiu-Yun Li ◽  
Yong-Heng Luo ◽  
En-Hua Xiao
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Hepatic Differentiation ◽  
Specific Factors

Sodium Butyrate Pre-Treatment Enhance Differentiation of Bone Marrow Mesenchymal Stem Cells (BM-MSCs) into Hepatocytes and Improve Liver Injury

2021 ◽  
Vol 21 ◽  
Author(s):  
Qiu-Yun Li ◽  
Juan Chen ◽  
Yong-Heng Luo ◽  
Wei Zhang ◽  
En-Hua Xiao
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Dependent Manner ◽  
Hepatic Differentiation ◽  
Marrow Mesenchymal Stem Cells ◽  
Specific Factors ◽  
Pre Treatment ◽  
Dose Dependent

Objective: The treatment of liver failure by stem cell transplantation has attracted growing interest. Herein, we aim to explore the role of sodium butyrate (NaB) in the hepatic differentiation of bone marrow mesenchymal stem cells (BM-MSCs) under liver-specific factors induction in vitro and vivo. Materials & Methods: We isolated BM-MSCs from the mononuclear cell fraction of rabbit bone marrow samples, and identified the cells by Immunophenotypic analysis. We investigated the effects of different concentrations and induction conditions. The histone deacetylase inhibitor NaB induced hepatic differentiation of BM-MSCs under liver-specific factors induction in vitro. Morphological features, liver-specific gene and protein expression, and functional analyses in vitro and vivo were performed to evaluate the hepatic differentiation of BM-MSCs. Results: Our results showed that pre-treated NaB inhibited the expression of liver-specific protein in a dose-dependent manner. The induction efficiency of NaB with 24h pre-treatment was higher than that of NaB continuous intervention. 0.5 mM 24h NaB pre-treated cells can improve liver tissue damage in vivo. And the liver ALB, AAT and the serum TP were significantly increased, while the serum ALT was significantly reduced. Conclusion: Continuous NaB treatment can inhibit BM-MSCs proliferation in a dose-dependent manner at a certain concentration range. 0.5 mM 24h pre-treatment of NaB enhanced differentiation of BM-MSCs into hepatocytes and improves liver injury in vitro and vivo.

In vitro hepatic differentiation of human bone marrow mesenchymal stem cells under differential exposure to liver-specific factors

2009 ◽  
Vol 154 (3) ◽  
pp. 122-132 ◽  
Author(s):  
Mihaela Chivu ◽  
Simona O. Dima ◽  
Cosmin I. Stancu ◽  
Camelia Dobrea ◽  
Valentina Uscatescu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Hepatic Differentiation ◽  
Marrow Mesenchymal Stem Cells ◽  
Specific Factors ◽  
Differential Exposure

296 IN VITRO DIFFERENTIATION OF PORCINE BONE MARROW-DERIVED MESENCHYMAL STEM CELLS INTO HEPATOCYTE-LIKE CELLS

2013 ◽  
Vol 25 (1) ◽  
pp. 295
Author(s):  
B. Mohana Kumar ◽  
W. J. Lee ◽  
Y. M. Lee ◽  
R. Patil ◽  
S. L. Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Class Ii ◽  
In Vitro Differentiation ◽  
Rt Pcr ◽  
Hepatic Differentiation ◽  
Alizarin Red ◽  
Differentiation Potential

Mesenchymal stem cells (MSC) are isolated from bone marrow or other tissues, and have properties of self renewal and multilineage differentiation ability. The current study investigated the in vitro differentiation potential of porcine bone marrow derived MSCs into hepatocyte-like cells. The MSC were isolated from the bone marrow of adult miniature pigs (7 months old, T-type, PWG Micro-pig®, PWG Genetics, Seoul, Korea) and adherent cells with fibroblast-like morphology were cultured on plastic. Isolated MSCs were positive for CD29, CD44, CD73, CD90, and vimentin, and negative for CD34, CD45, major histocompatibility complex-class II (MHC-class II), and swine leukocyte antigen-DR (SLA-DR) by flow cytometry analysis. Further, trilineage differentiation of MSC into osteocytes (alkaline phosphatase, von Kossa and Alizarin red), adipocytes (Oil Red O), and chondrocytes (Alcian blue) was confirmed. Differentiation of MSC into hepatocyte-like cells was induced with sequential supplementation of growth factors, cytokines, and hormones for 21 days as described previously (Taléns-Visconti et al. 2006 World J. Gastroenterol. 12, 5834–5845). Morphological analysis, expression of liver-specific markers, and functional assays were performed to evaluate the hepatic differentiation of MSC. Under hepatogenic conditions, MSC acquired cuboidal morphology with cytoplasmic granules. These hepatocyte-like cells expressed α-fetoprotein (AFP), albumin (ALB), cytokeratin 18 (CK18), cytochrome P450 7A1 (CYP7A1), and hepatocyte nuclear factor 1 (HNF-1) markers by immunofluorescence assay. In addition, the expression of selected markers was demonstrated by Western blotting analysis. In accordance with these features, RT-PCR revealed transcripts of AFP, ALB, CK18, CYP7A1, and HNF-1α. Further, the relative expression levels of these transcripts were analysed by quantitative RT-PCR after normalizing to the expression of the endogenous control, glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Data were analysed statistically by one-way ANOVA using PASW statistics 18 (SPSS Inc., Chicago, IL, USA), and significance was considered at P < 0.05. The results showed that the relative expressions of selected marker genes in hepatocyte-like cells were significantly increased compared with that in untreated MSC. The generated hepatocyte-like cells showed glycogen storage as analysed by periodic acid-Schiff (PAS) staining. Moreover, the induced cells produced urea at Day 21 of culture compared with control MSC. In conclusion, our results indicate the potential of porcine MSC to differentiate in vitro into hepatocyte-like cells. Further studies on the functional properties of hepatocyte-like cells are needed to use porcine MSC as an ideal source for liver cell therapy and preclinical drug evaluation. This work was supported by Basic Science Research Program through the National Research Foundation (NRF), funded by the Ministry of Education, Science and Technology (2010-0010528) and the Next-Generation BioGreen 21 Program (No. PJ009021), Rural Development Administration, Republic of Korea.

scholarly journals Zebularine Promotes Hepatic Differentiation of Rabbit Bone Marrow Mesenchymal Stem Cells by Interfering with p38 MAPK Signaling

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Yong-Heng Luo ◽  
Juan Chen ◽  
En-Hua Xiao ◽  
Qiu-Yun Li ◽  
Yong-Mei Luo
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Growth Factor ◽  
Hepatocyte Growth Factor ◽  
Hepatic Differentiation ◽  
Urea Production ◽  
Marrow Mesenchymal Stem Cells ◽  
Hepatocyte Growth

Demethylating agent zebularine is reported to be capable of inducing differentiation of stem cells by activation of methylated genes, though its function in hepatocyte differentiation is unclear. p38 signal pathway is involved in differentiation of hepatocytes and regulating of DNA methyltransferases 1 (DNMT1) expression. However, little is known about the impact of zebularine on bone marrow mesenchymal stem cells (BMMSCs) and p38 signaling during hepatic differentiation. The present study investigated the effects of zebularine on hepatic differentiation of rabbit BMMSCs, as well as the role of p38 on DNMT1 and hepatic differentiation, with the aim of developing a novel strategy for improving derivation of hepatocytes. BMMSCs were treated with zebularine at concentrations of 10, 20, 50, and 100 μM in the presence of hepatocyte growth factor; changes in the levels of hepatic-specific alpha-fetoprotein and albumin were detected and determined by RT-PCR, WB, and immunofluorescence staining. Expression of DNMT1 and phosphorylated p38 as well as urea production and ICG metabolism was also analyzed. Zebularine at concentrations of 10, 20, and 50 μM could not affect cell viability after 48 h. Zebularine treatment leads to an inhibition of DNMT activity and increase of hepatic-specific proteins alpha-fetoprotein and albumin in BMMSCs in vitro; zebularine addition also induced expression of urea production of and ICG metabolism. p38 signal was activated in BMMSCs simulated with HGF; inhibition of p38 facilitated the synthesis of DNMT1 and albumin in cells. Zebularine restrained DNMT1 and phosphorylated p38 which were induced by HGF. Therefore, this study demonstrated that treatment with zebularine exhibited terminal hepatic differentiation of BMMSCs in vitro in association with hepatocyte growth factor; p38 pathway at least partially participates in zebularine-induced hepatic differentiation of rabbit BMMSCs.

In vitro hepatic differentiation of human mesenchymal stem cells

Hepatology ◽  
2004 ◽  
Vol 40 (6) ◽  
pp. 1275-1284 ◽  
Author(s):  
Kuan-Der Lee ◽  
Tom Kwang-Chun Kuo ◽  
Jacqueline Whang-Peng ◽  
Yu-Fen Chung ◽  
Ching-Tai Lin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Human Mesenchymal Stem Cells ◽  
Hepatic Differentiation

In Vitro Hepatic Differentiation of Mesenchymal Stem Cells from Human Fetal Bone Marrow

2008 ◽  
Vol 36 (4) ◽  
pp. 721-727 ◽  
Author(s):  
X Wei ◽  
CY Wang ◽  
QP Liu ◽  
J Li ◽  
D Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Hepatic Differentiation ◽  
Fetal Bone

Cell Heterogeneity, Rather Than Cell Solvents Affects the Behavior of Mesenchymal Stem Cells in Vitro and Vivo

2020 ◽  
Author(s):  
Yong-Hong Wang ◽  
Ya-Chao Tao ◽  
Dong-Bo Wu ◽  
Meng-Lan Wang ◽  
Hong Tang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Survival Rates ◽  
Phosphate Buffer Solution ◽  
Human Umbilical Cord ◽  
Cell Heterogeneity ◽  
Hepatic Differentiation ◽  
Buffer Solution ◽  
Differentiation Potential

Abstract Background: Mesenchymal stem cells (MSCs) have been approved to treat various diseases, but they have to be expanded in vitro to gain sufficient cell doses. During the process of expansion, some obstacles remain to be addressed before MSCs translation to clinic. The purpose of this study was to investigate the effects of cell solvents and cell heterogeneity on the behavior of MSCs in vitro and vivo.Methods: Human umbilical cord MSCs (UC-MSCs) were dissolved in three different solvents: phosphate buffer solution (PBS), normal saline (NS) and dulbecco's modified eagle medium (DMEM). Their ultrastructure, viability and safety were explored and compared. MSCs from other two separate donors were grouped based on their mean diameters. The ultrastructure, proliferative and hepatic differentiation potential, senescent cell ratio and safety of the two UC-MSC aggregates were investigated and compared. The reason for mice death after UC-MSCs injection was further investigated.Results: The apoptosis rates, ultrastructure analysis and survival rates of mice among UC-MSCs in DMEM, NS and PBS were similar, and no significant differences were observed. The diameters of UC-MSCs of different sizes were measured. Cells with diameter of 15.58±3.813 μm were renamed as larger UC-MSC aggregates and cells with diameter of 19.14±4.885 μm were smaller aggregates. The mean diameter of larger MSC aggregates was significantly longer than that of smaller aggregates (p<0.01). Smaller MSCs had more potent proliferation potential and higher nucleus/ cytoplasm ratio than large ones. The number of cells positive for senescence-associated β-galactosidase staining was higher in larger UC-MSC aggregates. The survival rates of mice receiving 1×106 or 2×106 smaller MSCs were 100%, both higher than that receiving larger UC-MSCs sharing same amount. Meanwhile, the reason for mice death was explored and it revealed that larger UC-MSC aggregates were accumulated and evident in the pulmonary capillary lumen in dead mice.Conclusion: Solvents showed no significant effects on cell behavior, whereas, heterogeneity is quite prevalent in MSCs populations and may limit cell application, but it is easily overlooked. Hence it is necessary to establish a more precise standardization for culture-expanded MSCs and to improve MSCs manufacturing strategies.

Sign in / Sign up

Export Citation Format

Share Document