Differentiation of human umbilical cord mesenchymal stromal cells into low immunogenic hepatocyte-like cells.
OBJECTIVE
Mesenchymal stromal cells (MSC) isolated from several human tissues have been known to differentiate into the hepatic lineage in vitro, but the immunogenicity of the differentiated hepatocyte-like cells (DHC) has not been reported. Umbilical cord (UC) MSC are thought to be an attractive cell source for cell therapy because of their young age and low infection rate compared with adult tissue MSC.


METHODS
Hepatic differentiation of UC-MSC was induced with a 2-step protocol. The expressions of hepatic markers were detected by RT-PCR and immunofluorescence staining. Albumin production and urea secretion were measured by ELISA and colorimetric assay respectively. The immunosuppressive properties of DHC was detected by mixed lymphocyte culture.


RESULTS
After incubation with specific growth factors, including hepatocyte growth factor (HGF) and basic fibroblast growth factor (bFGF), UC MSC exhibited a high hepatic differentiation ability in an adherent culture condition. The differentiated UC MSC showed hepatocyte-like morphology and expressed several liver-specific markers at gene and protein levels. Furthermore, the DHC exhibited hepatocyte-specific functions, including albumin secretion, low-density lipoprotein uptake and urea production. More importantly, DHC did not express major histocompatibility complex (MHC) II antigen and were not able to induce lymphocyte proliferation in mixed lymphocyte culture, as undifferentiated UC MSC did.


CONCLUSIONS
Our results indicate that UC MSC are able to differentiate into functional hepatocyte-like cells that still retain their low immunogenicity in vitro. More importantly, DHC incorporated into the parenchyma of liver when transplanted into mice with CCl(4)-induced liver injury. Therefore, DHC may be an ideal source for cell therapy of liver diseases.
Zhao
Qinjun
Q
State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union of Medical College, Tianjin, China.
Ren
Hongying
H
Li
Xiyuan
X
Chen
Zhong
Z
Zhang
Xiangyu
X
Gong
Wei
W
Liu
Yongjun
Y
Pang
Tianxiang
T
Han
Zhong Chao
ZC
D000818:Animals; D002251:Carbon Tetrachloride; D002454:Cell Differentiation; D002469:Cell Separation; D048430:Cell Shape; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006684:HLA-DR Antigens / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000276:immunology* / Q000637:transplantation; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D016513:Mice, SCID; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology*; D014470:Umbilical Cord / Q000166:cytology*
D006684:HLA-DR Antigens; D008077:Lipoproteins, LDL; D002251:Carbon Tetrachloride