Advances in in vitro induction of hepatocyte-like cells and the application of small-molecule compounds in inducing hepatocyte-like cells / 临床肝胆病杂志

The induction of hepatocyte-like cells (HLCs) in vitro is one of the effective ways to obtain a large number of useful hepatocyte, and these HLCs can be used in disease modeling, drug design, and toxicological evaluation. At present, the induction of HLCs in vitro is mainly achieved by introducing exogenous transcription factors, cytokines or small-molecule compounds. Since small-molecule compounds have the advantages of structural diversity, controllable time and dose, and convenient and safe operation, scientists are devoted to screening out the small-molecule compounds to replace exogenous transcription factors and cytokines, and such compounds have a promising application prospect in the field of regenerative medicine. This article reviews the studies on the in vitro induction of HLCs from pluripotent stem cells and other adult stem cells and summarizes the application of small-molecule compounds in the in vitro induction of HLCs, in order to provide ideas and references for the in vitro induction of HLCs.

Induction of peripheral blood mononuclear cells to hepatocyte-like cells and preliminary study of cell response to injury under the effect of acetaminophen / 中华肝脏病杂志

Objective: To establish a method for the induction of peripheral blood mononuclear cells to hepatocyte-like cells, and preliminarily investigate cell response to injury under the effect of acetaminophen (APAP). Methods: The surface marker CD45 of peripheral blood mononuclear cells wase detected cells by using flow cytometry and immunofluorescence methods. The cellular morphology of induced hepatocyte-like cells was observed under an inverted microscope. Real-time fluorescent quantitative PCR (RT-PCR) was used to detect the expression level of hepatocyte-specific genes, such as cytochrome (CY) P1A2, CYP3A4, CYP2C9, albumin (ALB), alpha-fetoprotein (AFP), and hepatocyte nuclear factor (HNF)4α mRNA. Immunofluorescence method was used to detect intracellular hepatocyte markers AFP, HNF4α, and ALB expression at the protein level. Biochemical analyzer was used to detect hepatocyte-specific secretory functions of AFP, ALB, and urea. Luciferase chemiluminescence method was used to detect the activity of key drug metabolizing enzyme CYP3A4. Colorimetric assay was used to detect the effect of the drug acetaminophen on hepatocyte-like cells, and alanine aminotransferase (ALT) was used as an indicator of liver cell injury. The statistical differences between the data were compared with t-test and rank-sum test. Results: The positive expression rate of CD45 cell surface markers isolated from peripheral blood mononuclear cells was about 98%, and hepatocyte-like cell morphology changes appeared on 15th day of induction. Compared with isolated mononuclear cells, CYP1A2, CYP3A4, CYP2C9, ALB, AFP and HNF4α mRNA was markedly elevated. The expression level of AFP, ALB and HNF4α protein were equally increased, and the secretory function of AFP, ALB and urea were enhanced. Compared with primary hepatocytes, CYP1A2, CYP2C9, AFP, HNF4α mRNA, and CYP3A4 mRNA did not decrease. The expression levels of AFP, ALB, and HNF4α proteins in the cells did not decrease, and the secretory function of AFP, ALB, and urea did not decrease. In addition, the CYP3A4 enzyme activity produced by hepatocyte-like cells was similar to that of primary hepatocytes. Compared with hepatocyte-like cells incubated without APAP, hepatocyte-like cells incubated with APAP had higher ALT level. Under the effect of APAP, the ALT level of hepatocyte-like cells was higher than isolated mononuclear cells. Conclusion: Peripheral blood mononuclear cells can be induced into hepatocyte-like cells with partial characteristics of hepatocytes, including the activity of CYP3A4, a key enzyme of hepatocyte drug metabolism. Additionally, preliminarily ALT secretory features reflect the hepatocytes injury under the effect of acetaminophen.

Stem cell expansion and hepatic differentiation: Measurement of related indicators and detection methods / 中国组织工程研究

BACKGROUND: Stem cells have wide application prospects in tissue engineering, regenerative medicine, cell therapy and drug research. In recent years, the applied research, such as bioartificial liver, requires large-scale and high-quality stem cells by expansion, and final obtains hepatocytes (hepatocyte-like cells) by directional differentiation. Through this way, how to efficiently obtain the differentiated cells, with excellent consistency, powerful function and uniform expression of markers, are the key problems to be solved. OBJECTIVE: To review the detection indicators and detection methods related to the process of stem cell expansion and hepatic differentiation, summarize the indicators and methods that have been applied to online detection, discuss the limitations of some indicators and methods applied to online detection, and prospect the indicators and detection technologies expected to be applied to online detection in the future. METHODS: PubMed, Web of Science, Elsevier and CNKI databases were retrieved for the articles concerning stem cell expansion and hepatic differentiation published from 1990 to 2019. The literature related to stem cell expansion and hepatic differentiation was collected. The keywords were “stem cells; expansion; hepatic differentiation; monitoring; real-time online” in English and Chinese, respectively. A total of 93 articles were searched, and finally 53 eligible articles were selected and reviewed. RESULTS AND CONCLUSION: There are many detection indicators and detection methods for stem cell expansion and hepatic differentiation. Among them, the detection indicators of the expansion process are mainly related to the maintenance of pluripotency, metabolic activity, survival rate, and cancer transformation trend of stem cells. The process of hepatic differentiation varies with cell types, in which, the pluripotent stem cells and the mesodermal lineage adult pluripotent stem cells should differentiate into the endoderm-oriented cells and then the mature hepatocytes (hepatocyte-like cells), while the endodermal lineage adult pluripotent stem cells mainly composed of hepatic stem/progenitor cells can directly differentiate into mature hepatocytes (hepatocyte-like cells). The specific markers, survival rate and metabolic activity of cells in different stages of differentiation are the focus of detection. At present, although the detection methods based on large-scale biochemical detection and analysis equipment have high reliability, they face the problems of poor real-time performance, high cost and difficulties in integration and miniaturization. Future concerns are focused on the screening of key detection indicators, quantification of detection criteria and realization of automatic online detection during stem cell expansion and hepatic differentiation.

Efficient Isolation and Long-term Red Fluorescent Nanodia-mond Labeling of Umbilical Cord Mesenchymal Stem Cells for the Effective Differentiation into Hepatocyte-like Cells

Abstract Fluorescent nanodiamond (FND) has been used for long-term cell labeling and in vivo cell tracking because they have good at photostability and biocompatibility. In this study, we evaluate the effect of fluorescent nanodiamond labeling on in vitro culture and differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs) into hepatocyte-like cells (HLCs). For hepatic differentiation of hUCMSCs, cells were induced with human hepatocyte growth factor, nicotinamide and Dexamethasone. FND was supplied in two experimental groups with 20 μg/mL and 100 μg/mL in 2 hours. The cell was assessed for FND uptake by laser scan microscopy and flow cytometry methods. The effect of FND on hUCMSCs was evaluated by the cell viability and growth assays as well as the differentiation throughout of morphology alterations or gene expression of anfa-fetoprotein, albumin, and hepatocyte nuclear factor 4α. The results showed that the labeling of hUCMSCs is efficient and easy and there was significant cellular uptake of FND. We did not observe any negative impacts of FND to the cell viability and growth. FND can be utilized for the long-term labeling and tracking of hUCSCs and HLCs in vivo studies.

Galectin-3 induces differentiation of rat bone marrow mesenchymal stem cells into hepatocyte-like cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of galectin-3 in inducing the differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into hepatocyte- like cells and explore the involvement of the signaling pathways in the induced cell differentiation.</p><p><b>METHODS</b>The third passage of cultured rat femoral BMSCs were treated with 0.5 μg/mL galectin-3, 20 ng/mL hepatocyte growth factor (HGF) or both to induce their differentiation, with untreated rat BMSCs and hepatocytes as controls. At 7, 14, 21 and 28 days of induction, the cells were examined for morphological changes followed by glycogen staining, quantitative real-time PCR and Western blotting. Gene microarray technique was used to examine the mRNA expression profile of the BMSCs induced with galectin-3. The BMSCs were also induced with galectin-3 in combination with XMU-MP-1, a Hippo signaling pathway inhibitor, after which Western blotting was performed to detect the expressions of YAP, P-YAP, ALB, AFP and CK-18 in the cells.</p><p><b>RESULTS</b>The cells isolated from the femoral bone marrow of SD rats showed a consistent surface marker phenotype with the BMSCs. Induction with galectin-3, HGF, or both all resulted in gradual morphological changes of the BMSCs into hepatocyte-like cells, and the cells with a combined induction for 28 days showed the highest morphological similarity with hepatocytes. The cells induced with galectin-3, HGF, or their combination for 28 days all showed increased positivity rate of glycogen staining, which was the highest in the cells with combined induction ( < 0.05) without significant difference between the cells induced with galectin-3 and HGF alone ( > 0.05). Induction with galectin-3 and HGF alone both increased the expressions of AFP, ALB and CK-18 mRNAs in the cells, and their expression levels were similar between the cells at 28 days ( > 0.05). Galectin-3 and HGF did not show an interactive effect on the mRNA expressions of AFP (=0.236, =0.640) or ALB (=50.639, =0.000), but had a synergistic effect on CK-18 mRNA expression (=50.639, =0.000). The protein expressions of AFP, ALB and CK18 were also increased in the induced cells but not detected in the cells without induction. Gene microarray results revealed 27 up-regulated genes and 62 down-regulated genes in galectin-3-induced BMSCs involving TGF-β, PI3K-Akt and Hippo signal pathways. Induction with galectin-3 and galectin-3+XMU-MP-1 increased YAP expression in the cells, and galectin-3+XMU-MP-1 was more efficient to induce the differentiation of the BMSCs.</p><p><b>CONCLUSIONS</b>Galectin-3 can induce the differentiation of rat BMSCs into hepatocyte-like cells, and the combination with HGF increases the efficiency of induced differentiation of the cells. TGF-β, PI3K-Akt and Hippo pathways are involved in the induced differentiation of the BMSCs, and inhibiting Hippo pathway can improve the induction efficiency.</p>

Research advances in hepatocyte-like cells from human induced pluripotent stem cells and their application / 中华肝脏病杂志

Induced pluripotent stem cells (iPSCs) have the potential of proliferation and differentiation into a variety of somatic cells, including hepatocyte-like cells (HLCs). HLCs from human iPSCs (hiPSC-HLCs) have similar features and functions as primary hepatocytes and are used as an efficient in vitro model of hepatocytes, which brings hope to studies on liver diseases and drug hepatotoxicity evaluation. This article reviews the research advances in hiPSC-HLCs and their application in the fields of disease model, drug hepatotoxicity evaluation, and cell transplantation and discusses the future perspectives of the application of hiPSC-HLCs.

Role of different substrate elasticity in regulating rapid differentiation of HepaRG cells into Hepatocyte-like cells / 实用医学杂志

Objective To investigate the potential of substrate elasticity in regulating rapid differentiation of HepaRG cells into hepatocyte-like cells ,and further provide hepatocytes for bioartificial liver. Methods The substrate elasticity was divided into 4 groups. The expressions of albumin(ALB)were detected by albumin-green fluorescent protein-reporter system (ALB-GFP-reporter system) and Image J software;the cell morphology was observed by microscope and the amounts of cell were detected by cell Titer-Blue cell viability assay kit (alamar blue). Results The results of ALB showed that at the 4th hour,the expressions of ALB inside the HepaRG cells between 4s group and 8s group,16 s group and Glass group were not statistically different (t = 0.791,1.389, 2.481,P>0.05);at the 4th day,the expressions of 4s group had statistical differences in comparison with those of 16s group and Glass group(t = 12.41,12.52,P 0.05);at the 7th day,the expressions of 4s group were statistically different from those of 8s group,16s group and Glass group(t=3.266,6.725,8.005,P0.05). Conclusion Soft substrate can promote differentiation of HepaRG cells.

Comparison of the hepatic differentiation potential of human umbilical cord mesenchymal stem cells with bone marrow mesenchymal stem cells / 中华肝胆外科杂志

Objective To compare the hepatic differentiation potential of human umbilical cord mesenchymal stem cells (UC-MSCs) with bone marrow mesenchymal stem cells (BM-MSCs).Methods UC-MSCs and BM-MSCs derived from passage 3 were induced by IMDM supplemented with 20 μg/L HGF and 20 mg/L α-FGF.The medium was changed twice a week.The concentrations of albumin and urea nitrogen from cultural medium were measured to compare the differentiation ability of the two cells.We also examined the expression of hepatic related genes by real-time RT-PCR.Results UC-MSCs manifested significandy stronger proliferation potential than BM-MSCs.Both UC-MSCs and BM-MSCs could be induced into hepatocyte-like cells.The morphology of UC-MSCs tended to be more mature than BM-MSCs and they had more cytoplasmic granules.After 4 weeks,the levels of albumin and urea nitrogen from the cultural medium of the UC-MSCs group were significantly higher than the BM-MSCs group (P ＜ 0.05).Real-time PCR showed the expressions of four liver related genes CK18,G6P,HGF and ALB in the UC-MSCs group were significantly higher than the BM-MSCs group (P ＜ 0.05).Conclusion UC-MSCs had higher hepatic differentiation potential than BM-MSCs.Thus,UC-MSCs are more suitable than BM-MSCs for tissue engineering in the treatment of end-stage liver diseases.

Functional Comparison of Human Embryonic Stem Cells and Induced Pluripotent Stem Cells as Sources of Hepatocyte-Like Cells

Pluripotent stem cells can differentiate into many cell types including mature hepatocytes, and can be used in the development of new drugs, treatment of diseases, and in basic research. In this study, we established a protocol leading to efficient hepatic differentiation, and compared the capacity to differentiate into the hepatocyte lineage of human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). Optimal combinations of cytokines and growth factors were added to embryoid bodies produced by both types of cell. Differentiation of the cells was assessed with optical and electron microscopes, and hepatic-specific transcripts and proteins were detected by quantitative reverse transcription polymerase chain reaction and immunocytochemistry, respectively. Both types of embryoid body produced polygonal hepatocyte-like cells accompanied by time-dependent up regulation of genes for α-fetoprotein, albumin (ALB), asialoglycoprotein1, CK8, CK18, CK19, CYP1A2, and CYP3A4, which are expressed in fetal and adult hepatocytes. Both types of cell displayed functions characteristic of mature hepatocytes such as accumulation of glycogen, secretion of ALB, and uptake of indocyanine green. And these cells are transplanted into mouse model. Our findings indicate that hESCs and hiPSCs have similar abilities to differentiate into hepatocyte in vitro using the protocol developed here, and these cells are transplantable into damaged liver.

The differentiation of human umbilical mesenchymal stem cells into hepatocyte-like cells and related benefits for liver cirrhosis rats / 中华肝胆外科杂志

Objective To observe the effects of hepatocyte-like cells differentiated from human umbilical cord mesenchymal stem cells (HuMSCs) on the liver function of the rats with liver cirrhosis.Methods Carbon tetrachloride was used to prepare rat model of liver cirrhosis.Then the rats in the experimental group received portal vein injection of 1 ml differentiated hepatocyte-like cells (1 × 107) ; the mesenchymal stem cell (MSC) group was injected with the same volume and number of MSCs; the model group was injected with the same volume of saline (NS) ; the normal rats were treated as control group.After transplantation,the rat angular vein blood and liver tissue were obtained for testing.Results One week after transplantation,compared with the model group,levels of serum alanine aminotransferase (ALT),aspartate aminotransferase (AST) and total bilirubin (TBil) in the experimental group significantly decreased (P ＜0.05),while the albumin (Alb) level increased significantly (P ＜0.05).Compared with the MSC group,the level of Alb in the experimental group also significantly increased (P ＜ 0.05),but there were no differences between the two groups of ALT,AST and TBil.4 weeks after transplantation,compared with the model group,levels of serum ALT,AST and TBil in the experimental group also significantly decreased (P ＜ 0.05),while Alb level increased significantly (P ＜ 0.05).Compared with the MSC group,the differences of the levels of Alb,ALT,AST and TBil were all statistically significant (P ＜ 0.05).Real-time PCR test results showed that the expressions of four liver-related genes of the MSC group and experimental group significantly increased comparing with the model group (P ＜ 0.05).And the experimental group showed higher expression level comparing with the MSC group (P ＜ 0.05).Conclusion The differentiated hepatocyte-like cells could improve hepatic function of patients with liver cirrhosis to a certain degree and showed greater advantage than MSC.

Concise Review: Differentiation of Human Adult Stem Cells Into Hepatocyte-like Cells In vitro

Adult stem cells (ASCs) are undifferentiated cells found throughout the body that divide to replenish dying cells and regenerate damaged tissues, which are the powerful sources for cell therapy and tissue engineering. Bone marrow-derived mesenchymal stem cells (BMSCs), adipose tissue-derived mesenchymal stem cells (ADSCs), and peripheral blood monocytes (PBMCs) are the common ASCs, and many studies indicated that ASCs isolated from various adult tissues could be induced to hepatocyte-like cells in vitro. However, the isolation, culture protocols, characterization of ASCs and hepatocyte-like cells are different. This review aims to describe the isolation and culture procedures for ASCs, to summarize the molecular characterization of ASCs, to characterize function of hepatocyte-like cells, and to discuss the future role of ASCs in cell therapy and tissue engineering.

Effects of intracellular magnetic labeling of stem cells with superparamagnetic iron oxide on the differentiation capability into hepatocyte-like cells / 国际生物医学工程杂志

Objective To investigate the effects of intracellular magnetic labeling of stem cells with superparamagnetic iron oxide (SPIO) on the cell differentiation capability into hepatocyte-like cells.Methods Adiposederived stem cells (ADSCs) were obtained from the inguinal fat tissue of Sprague-Dawley rats.ADSCs were labeled with co-incubation of poly-l-lysine (PLL) and SPIO (25 μg/mL).Intracellular iron uptake was analyzed qualitatively with light microscopy.The viability of ADSCs was evaluated with trypan blue staining.SPIO-labeled and unlabled ADSCs were subjected to differentiate into hepatocyte-like cells with hepatocyte growth factor (HGF).Liver marker gene such as albumin (ALB) was analyzed by RT-PCR.The cell viability between the labeled cell group and unlabeled cell group adopted two independent sample t-Test.Results Light microscopy results revealed intracytoplasmic iron uptake and nearly 100％ of cell labeling efficiency.SPIO-labeled ADSCs had an unaltered viability as compared with unlabeled ADSCs (P＞0.05).After induction,glycogen storage within cytoplasm can be found in the two group on 14 d and 21 d,and the cells with positive staining increased on day 21.The two groups both express the ALB on 14 d and 21 d,which expressed higher on 21 d.Conclusion Intracellular magnetic labeling with SPIO did not affect the viability and capability of ADSCs to differentiate into hepatocyte-like cells.

Establishment of the limited cell line of skin fibroblast and their differentiation to hepatic cells / 国际生物医学工程杂志

Objective To establish the long-term culture system for fetal skin fibroblast by performing long time in vitro cultivation of the cells,and study the potential of its differentiation to hepatocytes.Methods Fibroblast was isolated from human fetus skin tissue.Surface phenotypes of cells were detected by ICC and FCM,and biological characteristics were analyzed by the karyotype analysis and soft agar colony formation observation.ALB、CK18、CK19 were detected by ICC,glycogen stain by PAS,AFP and ALB mRNA by RT-PCR after P3～30cells were induced differentiation by cytokines of HGF,FGF4 and OSM.Results CD29,CD49f,HLA- Ⅰ and CD 105 were highly expressed while CD90 hardly in skin fibroblast.The rate of induced differentiation of fibroblast into hepatocyte-like cells was approximately 5％.The cells could be cultured in vitro for almost 50 passages with normal karyotype and no oncogenic and immortalized characteristics.Conclsion The skin fibroblast possesses the characteristic of mesenchymal stem cell and can be induced into hepatocyte-like cell in vitro.

Bone Marrow Mesenchymal Stem Cells Differentiated into Hepatocyte-like Cells Induced by Rebirth Liver Tissue Lixivium of Mouse in Vitro / 中国康复理论与实践

@#Objective To induce bone marrow mesenchymal stem cells (BMSCs) differentiated into hepatocyte-like cells with rebirth liver tissue lixivium of mouse in vitro.Methods Mouse BMSCs were isolated and directionally induced with rebirth liver tissue lixivium of mouse 36 h after partial hepatectomy (PH). The morphology of cells was observed under an invert microscope, and the characteristics of differentiated cells was identified by immunofluorescence test.Results 7 days after induced by liver lixivium, the spindle shaped BMSCs became round and resembled hepatocyte-like cells. 1～2 weeks later, the differentiated cells expressed albumin, CK8 and CK18, which were known as characteristic markers of the hepatocyte.Conclusion BMSCs of mouse can be differentiated into hepatocyte-like cells under induction of liver tissue lixivium of mouse.

Bone marrow mesenchymal stem cells from rats with hepatic fibrosis differentiate into hepatocyte-like cells in vitro / 基础医学与临床

Objective To investigate the difference of differentiation from rat bone marrow mesenchymal stem cells(MSCs) from normal group and hepatic fibrosis model group into hepatocyte-like cells.Methods Wistar rats were randomly divided into two groups: normal group and hepatic fibrosis model group.The liver fibrosis was induced by CCL4.MSCs were isolated by combining gradient density centrifugation with plastic adherence.Pure MSCs were obtained by cultivation and passage.The cells then treated with HGF and FGF-4.Levels of AFP and albumin from supernatant were determined on day 15,21 and 27.On day 27,cells of induced and non-induced were collected,glycogen store of hepatocytes and the expressions of CK-18 and CK-19 were detected.Results The level of AFP in induced MSCs was higher on day 15,21,27,and reached the peak on day 21;there was no significant difference between induced and non-induced MSCs in albumin levels on day 15,but on day 21,27,compared with the non-induced MSCs,the albumin level in the induced MSCs was higher and reaches its peak on day 27;glycogen storage of induced MSCs was measured on day 27 as compared with non-induced MSCs;the induced MSCs expressed CK-18 and CK-19 while the non-induced MSCs did not.Compared by the levels of AFP and albumin,there was no significant difference in differentiation effect of MSCs between the normal group and hepatic fibrosis model group.Conclusion Rat MSCs of hepatic fibrosis model group could differentiate into hepatocyte-like cells with hepatic phenotype and biological function in the presence HGF and FGF-4.

Hepatocyte growth factor and epidermal growth factor induce mesenchymal stem cells isolated from patients with hepatocirrhosis into hepatocyte-like cells in vitro / 第三军医大学学报

Objective To explore the possibility of differentiation of mesenchymal stem cells isolated from patients with hepatocirrhosis into hepatocyte-like cells induced by EGF and HGF and to lay basis for transplanted autologous bone marrow MSCs in treatment of liver disease at terminal stage.Methods Bone marrow cells were obtained from volunteers with liver cirrhosis.MSCs were separated by density gradient centrifugation and were cultured through adhere culture.MSCs were cultured in DMEM medium with HGF,EGF,HGF+EGF or no growth factor.The phenotypes of MSCs were identified by flow cytometry,immunohistochemistry,and Albumin levels in culture supernatants were determined by ELISA.Results Growth and division of adherent cells obtained from the patients with hepatocirrhosis were good and the phenotypes of MSCs were CD29 positive and CD34 negative.The shape of MSCs changed from long fusiform to polygonal or round on 21th-28th days in grow factor induced groups.Immunocytochemical analysis for CK18 and AFP showed positive staining reaction for AFP on 7th day,for CK18 on 21st and 28th day in grow factor induced groups with MSCs-induced Alb production increasing in a time-dependent manner.No markers of hepatocyte linear cells were detected in no growth factor induction group.Conclusion Both HGF and EGF can induce mesenchymal stem cells to differentiate hepatocyte-like cells alone or coordinately.

Isolation and identification of human fetal bone-derived mesenchymal stem cells and their differentiation to hepatocyte like cells / 中国病理生理杂志

AIM: To separate and identify the mesenchymal stem cells (MSCs) from human fetal bone and to study their differentiation to hepatocyte like cells under the action of chemical induction.METHODS: The MSCs from human fetal bone were isolated and purified according to the different growth characteristic of attaching to the wall of cell culture flask. The cell cycle and surface markers of MSCs were identified using flow cytometry. The MSCs were pre-induced by adding DMSO, ?-Me and 5-aza for 24 h, then adding the inductive medium of H-DMEM and rh-HGF to induce their differentiation to hepatocyte like cells (HLCs). HLCs were identified by the typical morphological change and the expression of special protein with the method of immunocytochemistry. RESULTS: The MSCs derived from human fetal bone expressed adhesion molecules CD29+, CD44+, but not antigens of hematopoietic CD34, CD45, and not antigens related to GVHD, such as HLA-DR, CD80 and CD86. Exposure of these cells to above-mentioned inductive agents resulted in obvious morphological change and an increase in expression of AFP and ALB. CONCLUSION: The results suggest the existence of plentiful MSCs in human fetal bone. MSCs derived from human fetal bone can easily differentiate to HLCs, and they have a lower immunogenic nature, which may provide the ideal source for tissue engineering (bioartificial liver) for cellular therapeutics.