ABSTRACT
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.
Subject(s)
Adult , Animals , Humans , Mice , Cytochrome P-450 CYP1A2 , Cytochrome P-450 CYP3A , Cytokines , Embryoid Bodies , Glycogen , Hepatocytes , Human Embryonic Stem Cells , Immunohistochemistry , In Vitro Techniques , Indocyanine Green , Induced Pluripotent Stem Cells , Intercellular Signaling Peptides and Proteins , Liver , Pluripotent Stem Cells , Polymerase Chain Reaction , Reverse Transcription , Up-RegulationABSTRACT
Objective To investigate the feasibility of differentiating human umbilical cord blood stem cells into hepatocytes. Methods Thirty-six BALB/c nude mice were randomly divided into experimental group and control group(18 in each of the group), and experimental group was again randomly divided into group A, B and C (six in each of the group). The mice in experimental group and control group were exposed to 350 cGy radiation produced by 60Co. After 3 h, karyocytes at different concentrations in the fresh human umbilical cord blood were injected into the mice in experimental group A, B, C via their tail veins, and the equal volume of normal sodium (NS) was also injected into control group via tail veins. After one month, carbon tetrachloride (CCl4) was injected into experimental group A, B and control group via abdominal cavity, and the equal volume of normal sodium was injected into experimental group C. After two months, immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR) were used to detect the expressions of human cytokeratin-18 (CK18), cytokeratin-19 (CK19) and albumin (ALB) in liver tissues of all mice. Results The expressions of CK18, CK19 and ALB in injured liver tissues were all positive, and the expressions of experimental group B were higher than those of experimental group A (P